Researchers from the Oregon Stem Cell Center in Portland, Oregon have demonstrated the existence of at least four separate subtypes of human insulin-producing beta cells that may be important in the understanding and treatment of diabetes.
“This study marks the first description of several different kinds of human insulin producing beta cells,” said Markus Grompe of the Oregon Stem Cell Center at Oregon Health Science University. “Some of the cells are better at releasing insulin than others, whereas others may regenerate quicker. Therefore, it is possible that people with different percentages of the subtypes are more prone to diabetes. Further understanding of cell characteristics could be the key to uncovering new treatment options, as well as the reason why some people are diabetic and others are not.”
Diabetes mellitus affects more than 29 million people in the United States. There are two main types of diabetes mellitus, type I and type II. Type I diabetes mellitus is caused by insufficient production of insulin. Type II diabetes mellitus is caused by insulin resistance or the inability of the body to properly respond to the insulin produced by the body. Type I diabetes mellitus results from dysfunction or loss of insulin producing beta cells in the endocrine portion of the pancreas. Insulin is a hormone that helps the body keep normal blood sugar levels, and incorporate sugar in the bloodstream into cells to grow and repair tissues. Previously, only a single variety of beta cell was known to exist. However, using human pancreatic islets, or clusters of up to 4,000 cells, Grompe and colleagues identified a method to identify and isolate four distinct types of beta cells. They also found that hundreds of genes were differently expressed between cell subtypes and that these distinct beta cell subtypes produced different amounts of insulin.
All type 2 diabetics had abnormal percentages of the subtypes, suggesting a possible role in the disease process. Additional research is needed to determine how different forms of diabetes – and other diseases – affect the new cell subtypes, as well as how researchers may take advantage of these differences for medical treatment.
A research team from the University of Iowa has designed a protocol that can create insulin-producing cells that help normalize blood-sugar levels in diabetic mice from skin cells. This discovery represents one of the first steps toward developing patient-specific cell replacement therapy for Type 1 diabetes. This research, which was led by Nicholas Zavazava from the department of internal medicine, was published in the journal PLoS ONE.
Zavazava and his coworker used human skin cells taken from punch biopsies and reprogrammed them to into induced pluripotent stem cells. These induced pluripotent stem cells were then differentiated in culture into pancreatic insulin-producing beta cells.
In culture, Zavazava’s cell made insulin in response to increased concentrations, but when they were implanted into diabetic mice, these cells responded to glucose, secreted insulin and worked to lower the blood-sugar levels in the mice to normal or near-normal levels.
Mind you, these induced pluripotent stem cell-derived beta cells were not as effective as pancreatic cells in controlling blood sugar levels, according to Zavazava in a UI news release. However, Zavazava and his team views the cells’ response in mice as an “encouraging first step” toward the goal of generating effective insulin-producing cells that potentially could be used to not just treat but cure Type 1 diabetes in humans.
“This raises the possibility that we could treat patients with diabetes with their own cells,” Zavazava said. “That would be a major advance, which will accelerate treatment of diabetes.”
Zavazava is also a member of UI’s Fraternal Order of Eagles Diabetes Research Center. This center is one of several groups whose aim is to create an alternative source of insulin-producing cells that can replace the pancreatic beta cells that die off in people with Type 1 diabetes.
According to the UI news release, this study is the first to use human induced pluripotent stem cells instead of embryonic stem cells to generate insulin-producing pancreatic beta cells. This protocol has the advantage of creating beta cells from a patient’s own cells include. This would eliminate the need to wait for a donor pancreas, since pancreas transplants are an option for treating Type 1 diabetes, but the demand for transplants is much greater than the availability of organs from deceased donors. The use of induced pluripotent stem cells would also eliminate the need for transplant patients to take immunosuppressive drugs. Finally, the use of induced pluripotent stem cells would also avoid the ethical concerns with treatments based on embryonic stem cells.
Diabetes researchers at Saint Louis University have discovered a way to prevent the onset of Type I diabetes mellitus in diabetic mice. This strategy involves inhibiting the autoimmune processes that result in the destruction of the insulin-secreting pancreatic beta cells.
Type I diabetes is a life-long disease that results from insufficient production of the vital anabolic hormone insulin. In most cases of Type I diabetes mellitus, the body’s immune system destroys insulin-producing beta cells, and this insulin deficiency causes high blood sugar levels, also known as hyperglycemia. Treatments for the disease require daily injections of insulin.
Dr. Thomas Burris, chair of the university’s pharmacological and physiological science department, and his colleagues, have published their results in the journal Endocrinology. IN this paper, they report a procedure that could potentially prevent the onset of the disease rather than just treating the symptoms
“None of the animals on the treatment developed diabetes even when we started treatment after significant beta cell damage had already occurred,” Burris explained in a prepared statement. “We believe this type of treatment would slow the progression of type I diabetes in people or potentially even eliminate the need for insulin therapy.”
A group of immune cells known as lymphocytes come in two main forms: B lymphocytes, which secrete the antibodies that bind to foreign cells and neutralize them, and T cells, which recognize foreign substances and regulate the immune response. There are several different types of T lymphocytes, but for the purposes of this discussion, two specific subtypes of T lymphocytes seem to be responsible for the onset of Type I diabetes. T “helper cells” that have the CD4 protein on their surfaces, and T “cytotoxic “ cells have the CD8 protein on their cell surfaces seem to play a role in the onset of Type I diabetes, but a third subtype of T lymphocyte has remained a bit of an enigma for some time. This subtype of T lymphocytes is a subcategory of CD4 T cells and secretes a protein called “interleukin 17,” and is, therefore, known as TH17.
Dr. Burris and his collaborators from the Department of Molecular Therapeutics at the Scripps Research Institute have been examining TH17 cells for some time and they came upon a pair of nuclear receptors that play a crucial role in the development of TH17 cells. Could hamstringing the maturation of TH17 cells delay the onset of Type 1 diabetes mellitus?
Burris and others targeted these receptors by using drugs that bound to them and prevented them from working. This prevented the maturation of the TH17 T lymphocytes. When two nuclear receptors, Retinoid-related orphan receptors alpha (ROR-alpha) and Gamma-t (ROR-gamma-t) were inhibited, they prevented the autoimmune response that destroyed the beta cells.
To block these ROR alpha and gamma t receptors, Burris and others used a selective ROR alpha inhibitor and a gamma t inverse agonist called SR1001 that was developed by Dr. Burris. These drugs significantly reduced diabetes in the mice that were treated with it.
These findings show that TH17 cells play a significant role in the onset of Type I diabetes, and suggest that the use of drugs like these that target this cell type may offer a new treatment for the illness.
According to the American Diabetes Association, only 5% of people with diabetes have the Type I form of the disease, which was previously known as juvenile diabetes because it is usually diagnosed in children and young adults. The organization said that over one-third of all research they conduct is dedicated to projects relevant to type 1 diabetes.
By switching off a single gene, Columbia Medical Center scientists have converted cells from the digestive tract into insulin-secreting cells. This suggests that drug treatments might be able to convert gut cells into insulin-secreting cells.
Senior author Domenico Accili said this of this work: “People have been talking about turning one cell into another for a long time, but until now we hadn’t gotten to the point of creating a fully functional insulin-producing cell by the manipulation of a single target.”
Accili’s work suggests that lost pancreatic beta cells might be replaced by retraining existing cells rather than transplanting new insulin-secreting cells. For nearly two decades, scientists have been trying to differentiate a wide variety of stem cells into pancreatic beta cells to treat type 1 diabetes. In type 1 diabetes, the patient’s insulin-producing beta cells are destroyed, usually by the patient’s own immune system. The patient becomes dependent on insulin shots in order to survive.
Without insulin, cells have no signal to take up sugar and metabolize it. Also muscles and the liver do not take up amino acids and make protein, and the body tends to waste away, ravaged by high blood sugar levels that progressively and relentlessly damage it without the means to repair this damage.
Insulin-producing beta cells can be made in the lab from several different types of stem cells, but the resulting beta cells often do not possess all the properties of naturally occurring beta cells.
This led Accili and others to attempt to transform existing cells into insulin-secreting beta cells. In previous work, Accili and others demonstrated that mouse intestinal cells could be converted into insulin-secreting cells (see Talchai C, et al., Nat Genet. 2012 44(4):406-12), This recent paper demonstrates that a similar technique also works in human intestinal cells.
The gene of interest, FOXO1, is indeed present in human gut endocrine progenitor and serotonin-producing cells. In order to determine in FOXO1 inhibition could induce the formation of insulin-secreting cells, Accili and others used human induced pluripotent stem cells (iPSCs) and small “gut organoids,” which are small balls of gut tissue that grow in culture.
Inhibition of FOXO1 by either introducing a mutant version of the gene that encoded a protein that soaked up all the wild-type protein or by using viruses that forced the expression of a small RNA that prevented the expression of the FOXO1 gene caused loss of FOXO1 activity. FOXO1 inhibition promoted the generation of insulin-positive cells within the gut organoids that express all the genes and proteins normally found in mature pancreatic β-cells. These transdifferentiated cells also released “C-peptide,” which is a byproduct of insulin production, in response to drugs that drive insulin secretion (insulin secretagogues). Furthermore, these cultured insulin-secreting cells and survive when transplanted into mice where they continue to secrete insulin in response to increased blood sugar concentrations.
The findings of Accili and his colleagues provide some evidence that gut-targeted FOXO1 inhibition or transplantation of cultured gut organoids made from iPSCs could serve as a source of insulin-producing cells to treat human diabetes.
This is a remarkable piece of research, but there is one thing that troubles me about it. If the patient’s immune system has been sensitized to beta cells, making new beta cells will simply give the immune system something else to attack. It seems to me that retraining to immune system needs to be done first before replacement of the beta cells can ever hope to succeed.
A research group from the Sanford-Burnham Medical Research Institute in La Jolla, San Diego, California has used pluripotent stem cells to make insulin-secreting pancreatic beta cells that are encapsulated in a porous capsule from which they secrete insulin in response to rising blood glucose levels.
“Our study critically evaluates some of the potential pitfalls of using stem cells to treat insulin-dependent diabetes,” said Pamela Itkin-Ansari, an adjunct assistant professor with a joint appointment at UC San Diego. “We have shown that encapsulated hESC-derived pancreatic cells are able to produce insulin in response to elevated glucose without an increase in the mass or their escape from the capsule. This means that the encapsulated cells are both fully functional and retrievable.”
For this particular study, Itkin-Ansari and her colleagues used glowing cells to ensure that their encapsulated cells stayed in the capsule. To encapsulate the cells, this group utilized a pouch-like encapsulation device made by TheraCyte, Inc. that features a bilaminar polytetrafluoroethylene (PTFE) membrane system. This pouch surrounds the cells and protects from the immune system of the host while giving cells access to nutrients and oxygen.
With respect to the cells, making insulin-secreting beta cells from embryonic stem cell lines have met with formidable challenges. Not only are beta cells differentiated from embryonic stem cells poorly functional, but upon transplantation, they tend to be fragile and poorly viable.
To circumvent this problem, encapsulation technology was tapped to protect donor cells from the ravages of the host immune system. However, an additional advance made by Itkin-Ansari and her colleagues is that when they encapsulated islet-precursor cells, derived from embryonic stem cells, these cells survived and differentiated into pancreatic beta cells. In fact, islet progenitor cells turn out to be the ideal cell type for encapsulation, since they are heartier, and differentiate into beta cells quite efficiently when encapsulated.
In their animal model tests, these cells remained encapsulated for up to 150 days. Also, as an added bonus, because the progenitor cells develop glucose responsiveness without significant changes in mass, they do not outgrow their capsules.
In order to properly get this protocol to work in humans, Itkin-Ansari and her group has to scale up the size of their capsules and the number of cells packaged into them. Another nagging question is, “How long will an implanted capsule last in a human patient?
“Given the goals and continued successful results, I expect to see the technology become a treatment option for patients with insulin-dependent diabetes,” said Itkin-Ansari.
To date, Itkin-Ansari and others have been able to successfully treat diabetic mice. The problem with these experiments is that they mice were made diabetic by treatment with a drug called beta-alloxan, which destroys the pancreatic beta cells. Human type 1 diabetic patients have an immune system that is sensitized to beta cells. Even though the encapsulation shields the beta cells from contact with the immune system, will this last in human patients with an aggressive immune response against their own beta cells? It seems to me that induced pluripotent cells made from the patient’s own cells would be a better choice in this case than an embryonic stem cell line.
Nevertheless, this is a fine piece of research for diabetic patients.
Type 1 diabetes mellitus results from destruction of insulin-producing beta cells in the pancreas. Diabetics have to give themselves routine shots of insulin. The hope that stem cells offer is the production of cells that can replace the lost beta cells. “We are looking for ways to make new beta cells for these patients to one day replace daily insulin injections,” says Ben Stanger, MD, PhD, assistant professor of Medicine in the Division of Gastroenterology, Perelman School of Medicine at the University of Pennsylvania.
Some diabetics have had beta cells from cadavers transplanted into their bodies to replace the missing beta cells. Such a procedure shows that replacement therapy is, in principle possible. Therefore, transplanting islet cells to restore normal blood sugar levels in type 1 diabetics could treat and even cure disease. Unfortunately, transplantable islet cells are in short supply, and stem cell-based approaches have a long way to go before they reach the clinic. However, Stanger and his colleagues have tried a different strategy for treating type 1 diabetes. “It’s a powerful idea that if you have the right combination of transcription factors you can make any cell into any other cell. It’s cellular alchemy,” comments Stanger.
New research from Stanger and a postdoctoral fellow in his laboratory, Yi-Ju Chen that was published in Cell Reports, describes the production of new insulin-making cells in the gut of laboratory animals by introducing three new transcription factors. This experiment raises the prospect of using directly reprogrammed adult cells as a source for new beta cells.
In 2008, Stanger and others in Doug Melton’s laboratory used three beta-cell reprogramming factors (Pdx1, MafA, and Ngn3, collectively called PMN) to convert pancreatic acinar cells (the cells in the pancreas that secrete enzymes rather than hormones) into cells that had many of the features of pancreatic beta cells.
Following this report, the Stanger and his team set out to determine if other cells types could be directly reprogrammed into beta cells. “We expressed PMN in a wide spectrum of tissues in one-to-two-month-old mice,” says Stanger. “Three days later the mice died of hypoglycemia.” It was clear that Stanger and his crew were on to something. Further work showed that some of the mouse cells were making way too much extra insulin and that killed the mice.
When the dead mice were autopsied, “we saw transient expression of the three factors in crypt cells of the intestine near the pancreas,” explained Stanger.
They dubbed these beta-like, transformed cells “neoislet” cells. These neoislet cells express insulin and show outward structural features akin to beta cells. These neoislets also respond to glucose and release insulin when exposed to glucose. The cells were also able to improve hyperglycemia in diabetic mice.
Stanger and his co-workers also figured out how to turn on the expression of PMN in only the intestinal crypt cells to prevent the deadly whole-body hypoglycemia side effect that first killed the mice.
In culture, the expression of PMN in human intestinal ‘‘organoids,’ which are miniature intestinal units grown in culture, also converted intestinal epithelial cells into beta-like cells.
“Our results demonstrate that the intestine could be an accessible and abundant source of functional insulin-producing cells,” says Stanger. “Our ultimate goal is to obtain epithelial cells from diabetes patients who have had endoscopies, expand these cells, add PMN to them to make beta-like cells, and then give them back to the patient as an alternate therapy. There is a long way to go for this to be possible, including improving the functional properties of the cells, so that they more closely resemble beta cells, and figuring out alternate ways of converting intestinal cells to beta-like cells without gene therapy.”
This is hopefully a grand start to what might be a cure for type 1 diabetes.
Diabetes mellitus results from an insufficiency of insulin (Type 1 diabetes) or an inability to properly respond to insulin (Type 2 diabetes). Type 1 diabetes is caused by an attack by the patient’s own immune system on their pancreatic beta cells, which synthesize and secrete insulin. It is a disease characterized by inflammation in the pancreas. This suggests that abatement of inflammation in the pancreas might provide relief and delay the onset of diabetes.
Mesenchymal stem cells isolated from umbilical cord connective tissue, which is also known as Wharton’s jelly (WJ-MSCs), have the ability to reverse inflammatory destruction and might provide a way to delay or even reverse the onset of Type 1 diabetes.
To test this possibility, Jianxia Hu, Yangang Wang, and their colleagues took 60 non-obese diabetic mice and divided them into four groups: a normal control group, a normal diabetic group, a WJ-MSCs prevention group that was treated with WJ-MSCs before the onset of diabetes, and a WJ-MSCs treatment group that was treated with WJ-MSCs after the onset of diabetes.
After their respective treatments, the onset time of diabetes, levels of fasting plasma glucose (FPG), fed blood glucose levels and C-peptide (an indication of the amount of insulin synthesized), regulation of cytokines, and islet cells were examined and evaluated.
After WJ-MSCs infusion, fasting and fed blood glucose levels in WJ-MSCs treatment group decreased to normal levels in 6-8 days and were maintained for 6 weeks. The levels of fasting C-peptide of the WJ-MSC-treated mice was higher compared to diabetic control mice. In the WJ-MSCs prevention group, WJ-MSCs protected mice from the onset of diabetes for 8-weeks, and the fasting C-peptide in this group was higher compared to the other two diabetic groups.
Other comparisons between the WJ-MSC-treated group and the diabetic control group, showed that levels of regulatory T-cells (that down-regulate autoinflammation), were high and levels of pro-inflammatory molecules such as IL-2, IFN-γ, and TNF-α. The degree of inflammation in the pancreas was also examined, and pancreatic inflammation was depressed, especially in the WJ-MSCs prevention group.
These experiments show that infusions of WJ-MSCs can down-regulate autoimmunity and facilitate the recovery of islet β-cells whether given before or after onset of Type 1 Diabetes Mellitus. THis suggests that WJ-MSCs might be an effective treatment for Type 1 Diabetes Mellitus.
Researchers from the University of Copenhagen, in collaboration with an international team of investigators, have successfully developed an innovative three-dimensional method to grow miniature pancreas from progenitor cells. The future goal of this research is to utilize this model system to fight against diabetes. This research was recently published in the journal Development.
The new method takes cell material from mice and grows them in vividly picturesque tree-like structures. The cells used were mouse embryonic pancreatic progenitors, and they were grown in a compound called Matrigel with accompanying cocktails of growth factors. In vitro maintenance and expansion of these pancreatic progenitors requires active Notch and FGF signaling, and therefore, this culture system recapitulated the in vivo conditions that give rise to the pancreas in the embryo.
Professor Anne Grapin-Botton and her team at the Danish Stem Cell Centre, in collaboration with colleagues from the Ecole Polytechnique Fédérale de Lausanne in Switzerland, have developed a three-dimensional culture method that takes pancreatic cells and vigorously expands them. This new method allows the cell material from mice to grow vividly into several distinct picturesque, tree-like structures. The method offers tremendous long-term potential in producing miniature human pancreas from human stem cells. Human miniature pancreas organoids would be valuable as models to test new drugs fast and effectively, without the use of animal models.
“The new method allows the cell material to take a three-dimensional shape enabling them to multiply more freely. It’s like a plant where you use effective fertilizer, think of the laboratory like a garden and the scientist being the gardener,” says Anne Grapin-Botton.
In culture, pancreatic cells neither thrive nor develop if they are alone. A minimum of four pancreatic cells, growing close together is required for these cells to undergo organoid development.
“We found that the cells of the pancreas develop better in a gel in three-dimensions than when they are attached and flattened at the bottom of a culture plate. Under optimal conditions, the initial clusters of a few cells have proliferated into 40,000 cells within a week. After growing a lot, they transform into cells that make either digestive enzymes or hormones like insulin and they self-organize into branched pancreatic organoids that are amazingly similar to the pancreas,” adds Anne Grapin-Botton.
The scientists used this system to discover that the cells of the pancreas are sensitive to their physical environment, and are influenced by such seemingly insignificant factors as the stiffness of the gel and contact with other cells.
An effective cellular therapy for diabetes is dependent on the production of sufficient quantities of functional beta-cells. Recent studies have enabled the production of pancreatic precursors but efforts to expand these cells and differentiate them into insulin-producing beta-cells have proved a challenge.
“We think this is an important step towards the production of cells for diabetes therapy, both to produce mini-organs for drug testing and insulin-producing cells as spare parts. We show that the pancreatic cells care not only about how you feed them but need to be grown in the right physical environment. We are now trying to adapt this method to human stem cells,” adds Anne Grapin-Botton.
There has been a robust debate as to whether or not the pancreas has a stem cell population. Several studies suggested that the pancreatic duct cells could differentiate into hormone-secreting pancreatic cells. Unfortunately, when the cells of the pancreatic duct are marked, they clearly never contribute to regeneration of the pancreas. According to an article that appeared in the journal Developmental Cell by Oren Ziv, Benjamin Glaser, and Yuval Dor entitled “The Plastic Pancreas,” tying off the pancreatic duct kills off the acinar cells, but it leads to a large increase in the number of hormone-secreting beta cells. Something seems to be contributing cells to the adult pancreas. However when lineage studies tried to confirm that the pancreatic duct cells formed the new cells, it failed to find any connection between the new cells in the pancreas and the duct.
Recent experiments from Chris Wright’s lab suggest that the acinar cells are a population of progenitor cells that divide and differentiate into different kinds of pancreatic cell types after injury to the pancreas. A similar result was observed in work by Desai and others. If that’s not odd enough for you, another set of experiments from Pedro Herrera research group has shown once all the insulin-secreting beta cells are killed off, the adjacent glucagon-secreting cells transdifferentiate into insulin-secreting beta cells. Therefore, something interesting is afoot in the pancreas.
All these experiments were done with rodents. Whether or not they are transferable to human remains uncertain. Nevertheless, a fascinating paper in EMBO Journal from Hans Clevers lab at the Hubrecht Institute, Utrecht, Netherlands haws succeeded in culturing pancreatic precursor cells.
Here’s how they did it. Clevers and his crew took the pancreatic duct of mice and partially tied it off. In order to stem cells from the digestive tract to grow, they must upregulate a signaling pathway called the “Wnt” pathway. The Wnt pathway is quiet in the pancreas, but one the pancreas is injured, the Wnt pathway swings into gear and the cells begin to divide.
When Clevers and company dropped pancreatic duct tissue into culture, Wnt signaling activity soared and the cells grew into a mini-organ (organoid) that resembled and tiny pancreas in a culture dish. In fact, a single cell taken from the pancreatic duct could be cultured into an organoid.
This experiment shows that there are techniques for growing unlimited quantities of pancreatic cells. The therapeutic possibilities of this technology is tremendous. In Clever’s own words, “We have found a way to activate the Wnt pathway to produce an unlimited expansion of pancreatic stem cells isolated from mice. By changing the growth conditions we can select two different fates for the stem cells and generate large numbers of either hormone-producing beta cells or pancreatic duct cells.”
Can this work with human pancreatic duct cells? That is the $64,000 question. Clevers and his groups will almost certainly try to answer this questions next. If Clevers and his crew can get this to work, then the possibilities are vast indeed.
A research group that is part of the New York Stem Cell Foundation or NYSCF has generated patient-specific beta cells (the cells that secrete insulin in the pancreas), and these cultured beta cells accurately recapitulate the features of maturity-onset diabetes of the young or MODY.
In this research, NYSCF scientists and researchers from the Naomi Berrie Diabetes Center of Columbia University used skin cells from MODY patients to produce induced pluripotent stem cells (iPSCs) that were differentiated in the culture dish to form insulin-secreting pancreatic beta cells (if that sounds like a lot of work that’s because it is).
Other laboratories have succeeded in generating beta cells from embryonic stem cells and iPSCs, but questions remain as to whether or not these cells accurately recapitulate genetically-acquired forms of diabetes mellitus.
Senior co-author of this study, Dieter Egli, a senior research fellow at NYSCF, said: “We focused on MODY, a form of diabetes that affects approximately one in 10,000 people. While patients and other models have yielded important clinical insights into this disease, we were particularly interested in its molecular aspects – how specific genes can affect responses to glucose by the beta cell.”
MODY is a genetically inherited form of diabetes mellitus, and the most common form of MODY, type 2, results from mutations in the glucokinase or GCK gene. Glucokinase is a liver-specific enzyme and it adds a phosphate group to sugar so that the sugar can be broken down to energy by means of a series of reactions known as “glycolysis.” Glucokinase catalyzes the first step of glycolysis in the liver and in pancreatic beta cells. Mutations in GCK increase the sugar concentration in order for GCK to properly metabolize sugar, and this increases blood sugar levels and increases the risk for vascular complications.
MODY patients are usually misdiagnosed as type 1 or type 2 diabetics, but proper diagnosis can greatly alter the treatment of this disease. Correctly diagnosing MODY can also alert family members that they too might be carriers or even susceptible to this disease.
NYSCF researchers worked with skin cells from two patients from the Berrie Center who had type 2 MODY. After reprogramming these skin cells to become iPSCs, they differentiated the cells into beta cells, These cells had the impaired GCK activity, but in order to compare them to something, the NYSCF group also made iPSCs with a genetically engineered version of GCK that was impaired in the same way as the GCK gene in these two patients, and another cell line with normal versions of the GCK gene. They used these iPSCs to make cultured beta cells.
“Our ability to create insulin-producing cells from skin cells, and then to manipulate the GCK gene in these cells using recently developed molecular methods, made it possible to definitely test several critical aspects of the utility of stem cells for the study of human disease,” said Haiqing Hua, lead author of this paper and a postdoctoral fellow in the Division of Molecular Genetics.
The beta cells made from these iPSCs were transplanted into mice and these mice were given an oral glucose tolerance test. An oral glucose tolerance test is used to diagnose diabetes mellitus. The patient fasts for 12 hours and then is given a concentrated glucose concentration (4 grams per kilogram body weight), which the patient drinks and then the blood glucose level is examined at 30-minute intervals. The blood glucose levels of diabetic patients will rise and only go down very sluggishly whereas the blood glucose levels of a nondiabetic patient will rise and then decrease as their pancreatic beta cells start to make insulin. Insulin signals cells to take up glucose and utilize it, which lowers the blood glucose levels. A reading of over 200 milligrams per deciliters
When mice with the transplanted beta cells made from iPSCs were given oral glucose tolerance tests, the beta cells from MODY patients showed decreased sensitivity to glucose. In other words, even in the presence of high blood sugar levels, the beta cells made from iPSCs that came from MODY patients secreted little insulin. However, high levels of amino acids, which are the precursors of proteins, also induces insulin secretion, and in this case, beta cells from MODY patients secreted sufficient quantities of insulin.
When the iPSCs made from cells taken from MODY patients were subjected to genetic engineering techniques that repaired the defect in the GCK gene, these iPSCs differentiated into beta cells that responded normally to high blood glucose levels and secreted insulin when the blood glucose levels rose.
By making beta cells from MODY patients and then correcting the genetic defect in them and returning them to normal glucose sensitivity, NYSCF scientists showed that this type of diagnosis could lead to cures for MODY patients.
“These studies provide a critical proof-of-principle that genetic characteristics of patient-specific insulin-producing cells can be recapitulated through use of stem cell techniques and advanced molecular biological manipulation. This opens up strategies for the development of new approaches to the understanding, treatment, and, ultimately, prevention of more common types of diabetes,” said Rudolph Leibel of the Columbia University Medical Center.
Douglas Melton’s laboratory at the Harvard University Stem Cell Institute in Cambridge, Massachusetts has discovered a liver hormone that stimulates the growth of insulin-secreting beta cells in the pancreas. This discovery could very well lead to new treatments for diabetes.
This hormone, betatrophin, was induced in mice by treating them with a peptide that binds to insulin receptors. The insulin-occupied insulin receptors were unable to bind insulin, and that caused the animals to be resistant to insulin. Under these conditions, the livers of these mice produced betatrophin, which caused the animals’ insulin-secreting pancreatic β cells to proliferate. Melton and others searched for genes that showed increased activity under these insulin-resistant conditions, which allowed Melton and colleagues to isolate and identify betatrophin.
According to Melton and his co-workers, “Transient expression of betatrophin in mouse liver significantly and specifically promotes pancreatic β cell proliferation, expands β cell mass, and improves glucose tolerance” (from the abstract of the paper).
Further experiments showed that when eight-week-old mice injected with betatrophin there was an average 17-fold rise in the proliferation of their insulin-secreting pancreatic β cells. Melton and others published these results in the journal Cell. Fortunately, betatrophin is also found in the human liver, according to Melton and others.
“It’s rare that one discovers a new hormone, and this one is interesting because it’s so specific,” says Melton. “It works only on β cells and it’s so robust and so potent.”
Pancreatic β cells replicate rapidly during embryonic and neonatal stages in both mice and humans, but beta cell growth decreases dramatically in adults. A decrease in the function of beta cells late in life is the main cause of type 2 diabetes. Type 2 diabetes is a metabolic disorder that affects more than 300 million people worldwide. In the United States alone, the two forms of diabetes — type 2 and type 1— account for US$176 billion in direct medical costs each year.
Melton hypothesized that injections of betatrophin once a month, or even once a year, could potentially induce enough activity in pancreatic β cells to provide the same level of blood-sugar control for people with type 2 diabetes as do daily insulin injections. According to Melton, betatrophin would cause fewer complications, since the body would make its own insulin. He also hopes that betatrophin will be able to help people with type 1 diabetes.
Matthias Hebrok, director of the University of California, San Francisco, Diabetes Center, says that the work “is a great advance”. “The findings are very interesting,” he says, although he would like to see the experiments repeated in older mice. “Do mice that are on their way to becoming diabetic at an advanced age truly have an increase in proliferative capacity upon treatment with betatrophin?” he asks. This is a fair question.
Henrik Semb, managing director of the Danish Stem Cell Center in Copenhagen, says that “the identification of a factor, betatrophin, that stimulates mouse β-cell replication with remarkable efficiency is a very important discovery, because it provides the starting point for further studies to elucidate the underlying mechanism of β-cell replication.”
β-cell replication has proved difficult to control in humans, but producing enough betatrophin for testing in human clinical trials will take about two years, according to Melton, who is also working to identify the hormone’s receptor and its mechanism of action.
Type 1 diabetics and severe type 2 diabetics show reduction of insulin secretion as a result of destruction of the specific cells in the pancreas that produce insulin. These cells, the so-called beta cells, suffer destruction from the patient’s immune system (type 1 diabetes) or from overwork (type 2 diabetes). The holy grail of diabetes treatment is the regeneration of lost beta cells.
Several reports have marshaled evidence that the pancreatic beta cells do regenerate, but the constant assault by the immune system eventually destroys all the beta cells. Other reports have argued that a stem cell population in the ductal system of the pancreas can replenish the beta cells. Thus, augmenting beta cell regeneration seemed to be simply a matter of employing the already-present regenerative properties of the pancreas.
Unfortunately, a recent study seems to put the kibosh on any hope that the pancreatic beta cells regenerate. This new study was published in the Journal of Clinical Investigation. In this paper, researchers at Children’s Hospital of Pittsburgh report were unable to find signs of new beta cell production in several common models of pancreatic injury (see Xiao, X., et al. 2013. No evidence for beta-cell neogenesis in murine adult pancreas. J Clin Invest., 123(5):2207-17).
“Overall, the paper puts one more nail in what was already becoming an increasingly tight coffin for what had been the prevailing hypothesis about β-cell neogenesis in adult mice,” said Fred Levine, who studies β-cell regeneration at Sanford Burnham Medical Research Institute in La Jolla, California and was not involved in the study. Still, Levine cautions that this negative result does not completely rule out adult regeneration of β-cells in other injury models.
To detect the formation of new beta cells, George Gittes and his colleagues used an old cell tracking method, but applied it in a different manner. They used two fluorescent tags in transgenic mice: a red tag that targets a protein in the cell membrane of most cells in the body, except for insulin producing cells, and a green tag that only tagged pancreatic beta cells. Gittes team looked for cells that turned on their insulin genes for the first time during a 40–48 hour window. The cells, therefore, would express both tags and, as a result, appeared yellow.
The yellow transition was detected in embryonic mice, where neogenesis (new beta cell production) is expected to occur. But when the researchers examined adult cells, they saw no yellow cells—meaning no evidence of neogenesis. They repeated this experiment in several common models of pancreatic damage. For example, the pancreatic duct ligation model (PDL damages other pancreatic cell types but not β-cells. The absence of detectable neogenesis in these models “puts pressure on us to find models in which there is neogenesis,” said Gittes. But he remains “very confident” that there are other models in which neogenesis occurs.
In fact, several models not tested in this paper have shown evidence of neogenesis, including one of Gittes’ own. In 2011, in which his team found evidence of neogenesis in a mice who beta cells were engineered to express diphtheria toxin receptors, that led to their death (see Criscimanna, A., et al. 2011. Duct cells contribute to regeneration of endocrine and acinar cells following pancreatic damage in adult mice. Gastroenterology, 141(4):1451-62). In 2010, two other research groups, including one headed by Levine, also demonstrated neogenesis in adult mice through trandifferentiation of preexisting α-cells in pancreatic islets into β-cells (Thorel, F., et al. 2010. Conversion of adult pancreatic alpha-cells to beta-cells after extreme beta-cell loss. Nature,464(7292):1149-54 and Chung, C.H., et al. 2010. Pancreatic β-cell neogenesis by direct conversion from mature α-cells. Stem Cells, 28(9):1630-8).
“Overall, I believe that the pathway by which β-cell regeneration occurs…is likely to vary depending on the stimulus for regeneration,” said Levine. Therefore, the current work does not rule out neogenesis, even from duct cells, in other models. “I would argue that the old cliché, ‘Absence of evidence is not evidence of absence’ should be kept in mind when evaluating studies like this.”
Type 1 diabetes results from an inability to produce sufficient quantities of the hormone insulin. Without insulin, the body does not receive the signal to take up sugar from the blood, and the result is high blood sugar levels, which are damaging to tissues, and a general wasting of tissues because they cannot take up enough sugar to feed them.
The cells in the pancreas that produce insulin are the beta cells, and animal studies have shown that transplantation of new beta cells into diabetic animals can reverse and even in some cases cure the diabetic animals. Therefore researchers have tried to make beta cells from pluripotent stem cells in order to make a source of beta cells for transplantation.
Unfortunately, beta cell production in the laboratory has been fraught with problems. The cells produced by differentiation of embryonic stem cells do not have the characteristics of mature beta cells and they produce little insulin and are not glucose responsive (D’Amour, et al., (2006) Nat Biotechnol 24, 1392-1401).
A different strategy, however, works much better. Instead of differentiating stem cells into beta cells, differentiate them into those cells that will form beta cells and other types of pancreatic cells in the embryo – immature endocrine cell precursors – and then transplant those into the pancreas of diabetic mice. In this case, the endocrine cell precursors differentiate in the bodies of the mice into pancreatic beta cells that greatly resemble normal beta cells.
Why don’t embryonic stem cells for beta cells in culture? This question was pursued by a collaboration between research team led by Maike Sander at UC San Diego and a company called ViaCyte, Inc.
When it comes to endocrine precursors transplanted into mice, Dr. Sander noted that, “We found that the endocrine cells retrieved from transplanted mice are remarkably similar to primary human endocrine cells.” He continued, “This shows that hESCs (human embryonic stem cells) can differentiate into endocrine cells that are almost indistinguishable from their primary human counterparts.”
Well, ESCs can make perfectly fine beta cells in the mouse body, but not in culture. What’s up with that?
Sander and her colleagues examined the gene expression patterns of embryonic stem cells as they were differentiated and compared them with the gene expression patterns in those cells that were transplanted into mice and allowed to differentiate inside the body of the mouse.
What Sander and her team found was astounding. As cells progress through their developmental program, particular genes are brought on-line and expressed, and then turned off as the cells passed through each stage of endocrine cell differentiation. The cellular machinery that shuts off genes after they have been activated consists of a family of proteins that remodel chromatin known as the Polycomb group (PcG). PcG-mediated repression of genes silenced those genes that were only turned on temporarily once they were no longer required.
In the transplanted endocrine precursors, Sanders and his team noted an orderly progression of genes that were turned on and then turned off once as needed. However, in the embryonic stem cells that were differentiated into beta cells in culture, they discovered that these cells failed to express the majority of genes critical for endocrine cell function. The main reason for this appeared to be that the PcG-mediated repression of genes was not fully eliminated when particular genes had to be expressed at specific developmental stages. Thus these cultured cells failed to fully eliminate PcG-mediated repression on endocrine-specific genes, which contributes to the abnormality of the culture-derived beta cells.
Sander commented: “This information will help devise protocols to generate functional insulin-producing beta cells in vitro. This will be important not only for cell therapies, but also for identifying disease mechanisms that underlie the pathogenesis of diabetes.”
Embryonic stem cell lines are made from four-five-day-old human embryos. At this stage of development, the embryo is a sphere of cells with two distinct cell populations; an outside layer of flat trophoblast cells and an inner clump of round inner cell mass (ICM) cells. The embryo consists of ~100 cells four days after fertilization, and ~150 cells five days after fertilization.
Embryonic stem cell (ESC) derivation involves the removal of the trophoblast cells (which are collectively called the trophectoderm) and the isolation of the ICM cells. There are several ways to remove the trophectoderm, but the most commonly-used technique is “immunosurgery,” which uses antibodies that bind to proteins on the surfaces of the trophectoderm, and serum to initiate destruction of the trophoblast cells. The isolated ICM cells are then cultured, and if they grow, they may produce an embryonic stem cell line.
Immunosurgery was first perfected by Davor Solter and Barbara B. Knowles on mouse embryos. They used an antiserum that was raised in rabbits when the rabbits were immunized against mouse spleen tissue. When mouse embryos were incubated with this antiserum plus serum from mice, all the cells of the mouse embryo died. However, if they used the rabbit antiserum and serum from guinea pig, then only the trophoblast cells were destroyed. For human embryonic stem cell derivation, the rabbits are immunized again human red blood cells, and this rabbit antiserum is used with guinea pig serum. The serum contains proteins called “complement,” which bind to cells that have antibodies attached to them and bore holes in those cells, thus destroying them.
When ICM cells are cultured, they are placed on a layer of mouse cells that have been treated with a chemical called mitomycin C to prevent them from dividing. These non-dividing cells act as “feeder cells” that keep the ICM cells from differentiating. Because ICM cells are grown on animal cells, they cannot be used for clinical purposes, since they will possess animal proteins can carbohydrates on their surfaces, which would be attacked by the patient’s immune system. However, several ESC lines have been derived without animal products, and it is possible to make ESC lines that would be safe or human use.
ESC derivation results in the destruction of human embryos. There is not two ways about it. Even though there are potential ways around this problem, the majority of ESC lines were made literally over the dead bodies of very young human beings. All the rationalization in the world (the embryo is too young, too small, too inchoate, too unwanted, going to die anyway, in the wrong place at the wrong time) do not undo the fact that the embryo is a very young human being, and making an ESC line from it ends his/her life.
Getting the ESC line to differentiate in what you want it to be is another problem. If any undifferentiated cells remain after differentiation, they can cause tumors. Therefore, there is a need to ensure that differentiation is efficient and complete. To this end, Doug Melton’s lab at Harvard University has published a remarkable paper in the journal Nature that uses mesenchymal stem cells from particular organs to direct the differentiation of ESC lines.
Melton’s lab, in particular Julie M. Sneddon and Malgorzata Borowiak (say that fast five times), established 16 lines of tissue-specific mesenchymal stem cells (MSCs) from embryonic, neonatal and adult mouse intestine, liver, spleen, and pancreas and human pancreas too. Then they cultured mouse ESCs on these MSC lines to determine if they could drive the ESCs to differentiate into pancreas cells. In the embryo, pancreatic precursors express several genes in a nested, hierarchical fashion. First, they express Sox17, which is a common endodermal marker, and then pancreatic progenitors all express Pdx1. Of these pancreatic progenitors, some express Ngn3 and these will become endocrine rather than exocrine cells, and othe the Ngn3-expressing cells, a few will become beta cells that make insulin.
Melton and his co-workers tried to determine if any of these genes was up-regulated in their ESC lines if that were co-cultured with their established MSC lines. They discovered that four lines – MSC1, 2, 3, & 4, all affected gene expression when co-cultured with ESCs. MSC 1 and 2 induced and increase in Sox17 expression and MSC 3 and 4 increased the expression of Ngn3 in ESCs.
These changes in gene expression were due to increased cell proliferation of cells actually expressing these genes and not due to differential survival. Also, no combination of growth factors could achieve the same results as the accompanying MSC lines. Thus there is more going on here than the MSCs just secreting the right growth factors. The MSCs must be making contact with the ESCs and inducing them to differentiate into a particular cell type.
Next Melton and his colleagues determined if this interaction with MSCs caused the ESCs to lose their ability to self-renew. The answer was a clear “no.” Even though these ESC lines were expressing genes characteristic of endodermal or pancreatic tissue, they did not lose their ability to differentiate into pancreatic tissue when appropriately induced to do so, and they also id not lose their ability to self-renew and grow competently in culture.
In a more stringent test, these ESCs that had been grown on tissue-specific MSCs were implanted into mice. As Melton points out in the paper, the “most efficient published protocols for in vitro differentiation of pluripotent cells to beta-cells yield only a small percentage (typically 0-15%) of insulin-positive cells, and these do not secrete insulin in a glucose-responsive manner.” Could the MSC-conditioned ESCs do any better?
Before implantation, the ESCs were differentiated into endodermal progenitors (Sox17-expressing cells), and co-cultured with MSCs for at least 3-7 passages. Then they were differentiated into beta cells and transplanted into mice. There were a few important controls that were used; Just saline, implantations of MSCs alone, and ESCs that had been differentiated into beta cells, but had never been passaged on MSCs. Finally, human pancreatic islets were used as a positive control.
The results were interesting to say the least. The saline and MSC alone implantations showed no insulin production with or without glucose. Likewise the human pancreatic islets made insulin in a glucose-dependent manner (no surprise there). The ESC-derived beta cells that had never been passaged on MSCs made insulin, and even showed some ability to respond to glucose and make more insulin after glucose ingestion. However, the beta cells derived from ESCs that had been passaged on MSCs made insulin in a glucose-dependent manner. The experiment produced a wide range of variability since the number of transplanted cells differed between each trial, but the implanted beta cells derived from ESCs passaged on tissue-specific MSCs definitely performed the best, and even did as well or better than the implanted human beta cells in some cases.
Melton notes at the end of his paper that this technique worked rather well for coaxing ESCs to form pancreatic derivatives, but it could very well be applicable to other systems as well. Also, it could probably work with induced pluripotent stem cells, which have many (though not all) of the characteristics of ESCs and can be made without killing human embryos. Thus another technique for increasing ESC differentiation seems to be on the table.
Insulin is a protein hormone made by the beta cells of the pancreatic islets. It signals to the liver, skeletal muscles, and fat tissue to take up glucose and store it as glycogen (a polymer of glucose), or to convert it into fat. Insulin also induces the uptake of amino acids by muscles and the liver to form protein. This makes insulin one of the most important anabolic (building) hormones in the body.
Without sufficient quantities of insulin, blood sugar levels soar, since cells do not have the signal to take up sugar. Large quantities of sugar are quite damaging to cells and tissues, and the accumulating damage causes blindness, kidney failure, heart failure, circulatory and peripheral nerve troubles and other ailments.
This pathological condition is known as diabetes mellitus, and treatment of it requires routine injections of insulin. In order to actually treat insulin, we must somehow replace the deleted or damaged beta cells. Stem care cell treatment can potentially do this, but the details are still being worked out.
Danish stem cell scientists have provided some insights into ways to convert stem cells into pancreatic beta cells. By examining pancreatic development in mice, Palle Serup and his research group discovered a new gene called “Mind Bomb-1” that plays a role in pancreatic beta cell formation.
Accord to Dr., Serup, “To get stem cells to develop into insulin-producing beta cells, it is necessary tp know what signaling mechanisms normally control the creation of beta cells during fetal development. This is what our new research results can contribute. When we know the signaling paths, we can copy then in test tubes and thus in time convert stem cells to beta cells.” Dr. Serup is a member of the Danish Stem Cell Center or DanStem at the University of Copenhagen.
In a collaboration with researchers at DanStem, the Danish Hagedorn Research Institute, and other international partners in Japan, Germany, South Korea and the United States, these new findings were published in the April edition of the Proceedings of the National Academy of Sciences.
Previous work has established that during the early hours of the development of the pancreas, a signaling pathway that utilizes the “Notch” protein prevents pancreatic cells from differentiating into endocrine (hormone-making) cells and promotes the continued growth and proliferation of a kind of generic, all-purpose pancreas precursor cell. These all-purpose pancreatic precursor cells are called multipotent progenitor cells or MPCs, and they express two genes: Nkx6-1, and Ptf1.
A bit later, Nkx6 and Ptf1a start to antagonize each other such that cells that express Nkx6 cannot express and Ptf1 and Ptf1-expressing cells cannot express Nkx6. This antagonism between these two genes segregates the developing pancreas into two domains. The bit that is furthest away from the ductal system expresses Ptf1a+ and form “acinar progenitors.” The acinar cells are the clusters that make all the digestive enzymes released by the pancreas the bicarbonate ions. The portion of the developing pancreas that is closet to the ductal system expresses Nkx6-1, and makes the pancreatic duct and β-cell progenitors (see Russ HA, Efrat S. Pediatr Endocrinol Rev. 2011 Dec;9(2):590-7).
This sounds simple, but there are still several gaps that have yet to be filled in. For example, the signals that regulate patterning of the incipient pancreas and cause the segregation of the cells from one end to the other. Also, what dictates the formation of β-cell progenitors as opposed to ductal cells is also presently unknown.
In this present article, Serup and his colleagues discovered that deleting Mind Bomb-1 activity from the developing pancreas preventing the segregation of MPCs into Nkx6-expressing and Ptf1a-expressing cells. Instead the Nkx6-1-expressing cells were replaced by Ptf1-expressing cells. This prevented the formation of beta cells.
Interestingly, Serup and his team found that once the Notch protein acts early during pancreatic development, it actually acts again to help establish the segregated pancreas with Nkx6-1-expressing cells at one end and Ptf1a-expressing cells at the other. This shows that Notch is not only necessary early on, but also later for beta cell formation.
According the Serup, “Our research contributes knowledge about the next step in development and the signaling involved in the communication between cells – an area that has not been extensively described. This new knowledge about the ability of the so-called “Notch” signaling first to inhibit and then to stimulate the creation of hormone-producing cells is crucially important to being able to control stem cells better when working with them in test tubes.”
Type 1 diabetes results from an inability to make sufficient quantities of insulin. Insulin is made by specific cells in the pancreatic islets (also known as the islet of Langerhans). Most type 1 diabetics have suffered destruction of their pancreatic beta cells. Beta cell destruction can result from physical trauma to the pancreas, which causes the digestive enzymes of the pancreas to destroy the beta cells. For example, pancreatitis, pancreatic surgery, or certain industrial chemicals can cause diabetes. Also, particular drugs can also cause temporary diabetes, such as corticosteroids, beta blockers, and phenytoin. Rare genetic disorders (Klinefelter syndrome, Huntington’s chorea, Wolfram syndrome, leprechaunism, Rabson-Mendenhall syndrome, lipoatrophic diabetes, and others) and hormonal disorders (acromegaly, Cushing syndrome, pheochromocytoma, hyperthyroidism, somatostatinoma, aldosteronoma) also increase the risk for diabetes.
Additionally, viral infections of pancreas can cause the immune response to destroy pancreatic cells, and this wipes out enough beta cells to cause the onset of type 1 diabetes. The coxsackievirus family of viruses is a family of enteric viruses that are cause infections that are sometimes associated with the onset of type 1 diabetes, as are mumps and congenital rubella. In most cases, genetic factors cause the immune system to view the pancreatic beta cells as foreign invaders, and the beta cells are attacked and destroyed. Researchers have found at least 18 genetic loci that are designated IDDM1 – IDDM18 that are related to type 1 diabetes. The IDDM1 region contains the “HLA genes” that encode proteins called “major histocompatibility complex”. HLA genes encode cell-surface proteins that act as “bar codes” for the immune system. When cells have the proper bar codes on their cell surfaces, the immune system recognizes those cells as being a part of the body in which they reside, and the immune system leaves them alone. Any cells that do not have the right bar codes are attacked and destroyed, which is known as the “graft versus host response.” Therefore, it is fair to say that HLA genes affect the immune response. New advances in genetic research are identifying other genetic components of type 1 diabetes. Other chromosomes and genes continue to be identified.
A recent paper attempts to cure type 1 diabetes by using umbilical cord stem cells. Umbilical cord stem cells (UCSCs) have the ability to greatly calm down the immune system. UCSCs secrete a wide variety of molecules that prevent immune cells from reacting to and destroying other cells, and also have many cell surface proteins that bind to the surfaces of immune cells and put them to sleep (see Abdi et al., Diabetes 2008;57:1759-67 & Aguayo-Mazzucato C. and Bonner-Weir S, Nature Reviews Endocrinology 2010;6:726-36).
Animal experiments have shown that co-culturing UCSCs with circulating immune cells alters the immune response against pancreatic beta cells and greatly increases the ability of the animal to regulate blood glucose levels (Zhao et al., PLoS ONE 2009;4:e4226). The UCSCs seem to “re-educate” the immune cells so that they do not recognize the pancreatic islets are foreign anymore. Therefore, Yong Zhao and his colleagues in Theodore Mazzone’s laboratory at the University of Chicago, IL, and collaborators at the General Hospital of Jinan Military Command, Shandong, China, used human UCSCs to re-educate immune cells in human type 1 diabetic patients. See here for this paper.
To do this, they circulated the blood of each patient through a close-loop system that separated the immune cells (lymphocytes) from whole blood. Thee lymphocytes were then co-cultured with UCSCs for 2-3 hours and then returned to the patients.
The results were remarkable. Six patients in group A, who all had some residual beta cell function showed successively improved insulin production 12-24 weeks after treatment. They also showed a reduced need for insulin shots, and overall improvement of their fasting blood glucose levels. Six patients in group B, who had no residual beta cell function, showed increased production of insulin production 12 week after treatment. This is an incredible finding because those without beta cells essentially grew new ones that were not attacked by the immune system. The group B group also saw successively reduced requirements for injected insulin. The patients in the control, whose immune cells did not undergo re-education by UCSCs showed no improvement.
Furthermore, the patients whose immune cells were re-educated by the UCSCs, did not experience any adverse effects. This procedure seems to be quite safe and feasible.
There is a word of caution here. These patients must be followed over several years to establish that the re-education of the lymphocytes is maintained over time. Also, this study is quite small and despite the amazing results, a larger study is needed. All the same, this is an incredible result that reverses type 1 diabetes, and even though caution is needed, embryonic stem cells were not required to do this.
Embryonic stem cells can be made from adult cells. Such cells are called iPSCs or induced embryonic stem cells, and they have all the characteristics of embryonic stem cells made by means of the destruction of embryos.
Lately, scientists have found a way to convert one type of adult cell into another type of adult without going through any embryonic step.
Qi Zhou and his colleagues from the Melton lab at Harvard were able to transform pancreatic enzyme-secreting cells (exocrine cells) into insulin-secreting cells by inserting three transcription factors (Ngn3, also known as Neurog3), Pdx1 and Mafa into the exocrine cells and they reprogrammed themselves into beta-cells (Nature 455, 627–32 (2008)). Also, Yechoor and his colleagues used a similar technique that placed neurogenin into liver cells in a live animal. These animals shows insulin-secreting cells into their livers, which showed that the liver cells had been reprogrammed into beta cells (V. Yechoor et al., Dev. Cell 16, 358–73 (2009)).
This shows that reprogramming is vastly superior and cheaper than making cloned embryos that we subsequently kill and use to make embryonic stem cells. This is the therapeutic way of the future.
Now, Jun Takeuchi and Benoit Bruneau at the Gladstone Institute of Cardiovascular Disease in San Francisco have found that adding cardiac-specific genes to developing mouse embryos can make even some extra-embryonic parts become beating heart cells. They made the cells from amnion, the thin layer that surrounds the embryo and fetus throughout development. This is the sac that breaks when we say that a mother’s water breaks. The amnion is normally medical waste, but can now be used to make heart cells.