Small Human Stomach Organoids Made From Induce Pluripotent Stem Cells


A new study published in the international journal Nature describes, for the first time, the use of human pluripotent stem cells to create a three-dimensional stomach-like mini-organ. This is the beginning of what might become an unprecedented tool for examining the genesis of diseases such as stomach cancer to diabetes.

Jim Wells and his colleagues at Cincinnati Children’s Hospital Medical Center used human pluripotent stem cells, which are made from mature human cells through a combination of genetic engineering and cell culture techniques, to grow a their miniature stomachs. Wells’ group then used their mini-stomachs also known as gastric organoids, in collaboration with scientists from University of Cincinnati College of Medicine, to study the infection of stomach tissue by the bacterium Helicobacter pylori, which causes peptic ulcer disease and stomach cancer.

According to Wells, a scientist in the divisions of Developmental Biology and Endocrinology at Cincinnati Children’s, this is the first time anyone has succeeded in making three-dimensional human gastric organoids (hGOs). This achievement may present new opportunities for drug discovery, modeling early stages of stomach cancer and studying some of the factors that give rise to obesity related diabetes. This work also represents the first time researchers have produced three-dimensional human embryonic foregut, which is a good starting point for generating other foregut organ tissues such as the lungs and pancreas. “Until this study, no one had generated gastric cells from human pluripotent stem cells (hPSCs),” Wells said. “In addition, we discovered how to promote formation of three-dimensional gastric tissue with complex architecture and cellular composition.”

a, Schematic representation of a typical antral gland showing normal cell types and associated molecular markers. b–g, Immunofluorescent staining demonstrated that day-34 hGOs consisted of normal cell types found in the antrum, but not the fundus. The hGO epithelium contained surface mucous cells that express MUC5AC (b, left), similar to the P12 mouse antrum (b, right), but not ATP4B-expressing parietal cells (c, left) that characterize the fundus (c, right). SOX9+ cells were found at the base of the hGO epithelium (d, left), similar to the progenitor cells found in the P12 antrum (d, right). Furthermore, hGOs contained MUC6+ antral gland cells (e) and LGR5-expressing cells (yellow arrow) (f). Boxed regions in b–f are shown as high magnification images below (b, c, d) or to the right (e, f) of the original. g, Day-34 hGOs also contained endocrine cells (SYP) that expressed the gastric hormones GAST, SST, GHRL and serotonin (5-HT). Scale bars, 100 μm (original images in b–f) and 20 μm (magnified images in b–f and g). Marker expression data are representative from a minimum of 10 independent experiments, except LGR5-eGFP data, which is a representative example from two separate experiments. DAPI, 4′,6-diamidine-2-phenylindole.

a, Schematic representation of a typical antral gland showing normal cell types and associated molecular markers. b–g, Immunofluorescent staining demonstrated that day-34 hGOs consisted of normal cell types found in the antrum, but not the fundus. The hGO epithelium contained surface mucous cells that express MUC5AC (b, left), similar to the P12 mouse antrum (b, right), but not ATP4B-expressing parietal cells (c, left) that characterize the fundus (c, right). SOX9+ cells were found at the base of the hGO epithelium (d, left), similar to the progenitor cells found in the P12 antrum (d, right). Furthermore, hGOs contained MUC6+ antral gland cells (e) and LGR5-expressing cells (yellow arrow) (f). Boxed regions in b–f are shown as high magnification images below (b, c, d) or to the right (e, f) of the original. g, Day-34 hGOs also contained endocrine cells (SYP) that expressed the gastric hormones GAST, SST, GHRL and serotonin (5-HT). Scale bars, 100 μm (original images in b–f) and 20 μm (magnified images in b–f and g). Marker expression data are representative from a minimum of 10 independent experiments, except LGR5-eGFP data, which is a representative example from two separate experiments. DAPI, 4′,6-diamidine-2-phenylindole.

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Wells’ gastric organoids are a significant advance in gastroenterological research because distinct differences in development and architecture of the adult stomach limit the reliability of mouse models for studying human stomach development and disease.

As a research tool, human gastric organoids may help clarify other unknown features of the stomach, such as identifying those biochemical processes in the gut that allow gastric-bypass patients to become diabetes-free soon after surgery before losing significant weight. Medical conditions such as obesity-fueled diabetes and the metabolic syndrome are of great interest to public health workers, given the explosion of global cases in the last few decades. A major challenge to addressing these and other medical conditions that involve the stomach has been a relative lack of reliable laboratory model systems to accurately recapitulate human biology.

The key to growing human gastric organoids was to identify the developmental steps involved in normal stomach formation. Manipulation of these processes in a cell culture system drove human pluripotent stem cells to form immature stomach tissue. In culture and over the course of a month, these steps resulted in the formation of 3D human gastric organoids that were around 3mm (1/10th of an inch) in diameter. Wells and his colleagues also used this approach to identify steps that go awry when the stomach does not form correctly.

In collaboration with his colleagues, Kyle McCracken, an MD/PhD graduate student in Wells’ laboratory, and Yana Zavros, PhD, a researcher at UC’s Department of Molecular and Cellular Physiology, Wells showed that his gastric organoids were rapidly infected by H. pylori bacteria. Within 24 hours of inoculation, the bacteria had triggered significant biochemical changes to the organ, and the human gastric organoids faithfully mimicked the early stages of H. pylori-induced gastric disease. McCracken also noticed activation of a cancer gene called c-Met, which is one of the first stages in the induction of stomach cancer, an important long-term sequel to peptic ulcer disease. McCracken was also surprised by the rapid spread of infection in the tissues of his human gastric organoids.

a, Day-34 hGOs contained a zone of MKI67+ proliferative cells similar to the embryonic (E18.5) and postnatal (P12) mouse antrum. b, Using hGOs to model human-specific disease processes of H. pylori infection. Pathogenic (G27) and attenuated (ΔCagA) bacteria were microinjected into the lumen of hGOs and after 24 h, bacteria (both G27 and ΔCagA strains) were tightly associated with the apical surface of the hGO epithelium. c, Immunoprecipitation (IP) for the oncogene c-Met demonstrates that H. pylori induced a robust activation (tyrosine phosphorylation (pTyr)) of c-Met, and this is a CagA-dependent process. Furthermore, CagA immunoprecipitated with c-Met, suggesting that these proteins interact in hGO epithelial cells. Phosphorylated c-Met (phos. c-MET) and CagA control lysates were not immunoprecipitated but used to confirm molecular masses. The molecular mass markers are indicated (130 and 170 kilodaltons (kDa)) and shown in Extended Data Fig. 9c. IB, immunoblotting. d, Within 24 h, H. pylori infection caused a CagA-dependent twofold increase in the number of proliferating cells in the hGO epithelium, measured by 5-ethynyl-2′-deoxyuridine (EdU) incorporation. *P < 0.05; two-tailed Student’s t-test; n = 3 biological replicates per condition, data representative of 4 independent experiments. Scale bars, 100 μm (a) and 20 μm (b). Error bars represent s.e.m.

a, Day-34 hGOs contained a zone of MKI67+ proliferative cells similar to the embryonic (E18.5) and postnatal (P12) mouse antrum. b, Using hGOs to model human-specific disease processes of H. pylori infection. Pathogenic (G27) and attenuated (ΔCagA) bacteria were microinjected into the lumen of hGOs and after 24 h, bacteria (both G27 and ΔCagA strains) were tightly associated with the apical surface of the hGO epithelium. c, Immunoprecipitation (IP) for the oncogene c-Met demonstrates that H. pylori induced a robust activation (tyrosine phosphorylation (pTyr)) of c-Met, and this is a CagA-dependent process. Furthermore, CagA immunoprecipitated with c-Met, suggesting that these proteins interact in hGO epithelial cells. Phosphorylated c-Met (phos. c-MET) and CagA control lysates were not immunoprecipitated but used to confirm molecular masses. The molecular mass markers are indicated (130 and 170 kilodaltons (kDa)) and shown in Extended Data Fig. 9c. IB, immunoblotting. d, Within 24 h, H. pylori infection caused a CagA-dependent twofold increase in the number of proliferating cells in the hGO epithelium, measured by 5-ethynyl-2′-deoxyuridine (EdU) incorporation. *P < 0.05; two-tailed Student’s t-test; n = 3 biological replicates per condition, data representative of 4 independent experiments. Scale bars, 100 μm (a) and 20 μm (b). Error bars represent s.e.m.

There is a relative dearth of literature on how the human stomach developments, which was a significant impediment to Wells’ research. Wells and his coworkers had to use a combination of published works and studies from his own lab, to answer a number of basic developmental questions about how the stomach forms. Over the course of two years, by experimenting with different factors to drive the formation of the stomach, Wells and his colleagues came upon a protocol that resulted in the formation of 3D human gastric tissues in culture.

a, Schematic representation of the in vitro culture system used to direct the differentiation of pluripotent stem cells into three-dimensional gastric organoids. b, Defining molecular domains of the posterior foregut in E10.5 mouse embryos with Sox2, Pdx1 and Cdx2; Sox2/Pdx1, antrum (a); Sox2, fundus (f); Pdx1, dorsal and ventral pancreas (dp and vp); Pdx1/Cdx2, duodenum (d). c, Posterior foregut spheroids exposed for three days to retinoic acid (2 μM) exhibited >100-fold induction of PDX1 compared to control spheroids, measured by qPCR at day 9. *P < 0.05; two-tailed Student’s t-test; n = 3 biological replicates per condition, data representative of 4 independent experiments. d, Time course qPCR analysis of antral differentiation (according to protocol detailed in Fig. 2a) demonstrated sequential activation of SOX2 at day 6 (posterior foregut (FG) endoderm), followed by induction of PDX1 at day 9 (presumptive antrum). Day-9 antral spheroids had a 500-fold increase in SOX2 and a 10,000-fold increase in PDX1 relative to day-3 definitive endoderm (DE). *P < 0.05; two-tailed Student’s t-test; n = 3 biological replicates per time point, data representative of 2 independent experiments. The pancreatic marker PTF1A was not significantly increased. e, Stereomicrographs showing morphological changes during growth of gastric organoids. By 4 weeks, the epithelium of hGOs exhibited a complex folded and glandular architecture (arrows). D, day. f, Comparison of mouse stomach at E18.5 and day-34 hGOs. Pdx1 was highly expressed in the mouse antrum but excluded from the fundus. Human gastric organoids expressed PDX1 throughout the epithelium and exhibited morphology similar to the late gestational mouse antrum (arrows). Scale bars, 100 μm (b and f) and 250 µm (e). Error bars represent s.d.

a, Schematic representation of the in vitro culture system used to direct the differentiation of pluripotent stem cells into three-dimensional gastric organoids. b, Defining molecular domains of the posterior foregut in E10.5 mouse embryos with Sox2, Pdx1 and Cdx2; Sox2/Pdx1, antrum (a); Sox2, fundus (f); Pdx1, dorsal and ventral pancreas (dp and vp); Pdx1/Cdx2, duodenum (d). c, Posterior foregut spheroids exposed for three days to retinoic acid (2 μM) exhibited >100-fold induction of PDX1 compared to control spheroids, measured by qPCR at day 9. *P < 0.05; two-tailed Student’s t-test; n = 3 biological replicates per condition, data representative of 4 independent experiments. d, Time course qPCR analysis of antral differentiation (according to protocol detailed in Fig. 2a) demonstrated sequential activation of SOX2 at day 6 (posterior foregut (FG) endoderm), followed by induction of PDX1 at day 9 (presumptive antrum). Day-9 antral spheroids had a 500-fold increase in SOX2 and a 10,000-fold increase in PDX1 relative to day-3 definitive endoderm (DE). *P < 0.05; two-tailed Student’s t-test; n = 3 biological replicates per time point, data representative of 2 independent experiments. The pancreatic marker PTF1A was not significantly increased. e, Stereomicrographs showing morphological changes during growth of gastric organoids. By 4 weeks, the epithelium of hGOs exhibited a complex folded and glandular architecture (arrows). D, day. f, Comparison of mouse stomach at E18.5 and day-34 hGOs. Pdx1 was highly expressed in the mouse antrum but excluded from the fundus. Human gastric organoids expressed PDX1 throughout the epithelium and exhibited morphology similar to the late gestational mouse antrum (arrows). Scale bars, 100 μm (b and f) and 250 µm (e). Error bars represent s.d.

Wells emphasized importance of basic research for the eventual success of this project, adding, “This milestone would not have been possible if it hadn’t been for previous studies from many other basic researchers on understanding embryonic organ development.”

While this does represent a terrific stride toward better model systems for gastric research and pathology, these gastric organoids are very immature and lack several of the cell types found in mature stomach tissue. For example, these organoids lack chief cells, which secrete the stomach enzyme pepsin (in an inactive form called pepsinogen), and parietal cells, which produce stomach acid. This is significant because chronic inflammation of the stomach can cause loss of parietal cells, which decreases chief cell differentiation and induce chief cells to transdifferentiate back into neck cells. This leads to overproduction of mucus cells. This mucus cell metaplasia is known as spasmolytic polypeptide expressing metaplasia (SPEM) that seems to be a precancerous condition for the stomach. Also if parietal cells are lost, mature chief cells do not form. This seems to imply that parietal cells secrete factors that lead to differentiation of chief cells, so if lost. These gastric organoids also do not make ECL cells or enterochromaffin-like cells, which secrete histamine, one of the most important regulators of stomach acid production. A prolonged stimulation of these ECL cells causes increased numbers of them. This is especially important in gastrinomas, which are tumors in which there is an excessive secretion of the stomach hormone gastrin, one of the key factors contributing to Zollinger-Ellison syndrome.  The hallmark of this disease is ulceration of the stomach and upper small intestine (duodenum) as a result of excessive and unregulated secretion of gastric acid.  Most commonly, hypergastrinemia is the result of these gastrin-secreting tumors or gastrinomas that develop in the pancreas or duodenum.  Thus, in only this short discussion, we have noted several diseases of the stomach that cannot be modeled with this particular system because these stomach-specific cells are not present.

Therefore, while this is a fantastic model system for stomach development and H. pylori infection, more work remains in order to make a stomach model that more accurately models the adult stomach.

“In Body” Muscle Regeneration


Researchers at Wake Forest Baptist Medical Center’s Institute for Regenerative Medicine have hit upon a new strategy for tissue healing: mobilizing the body’s stem cells to the site of injury. Thus harnessing the body’s natural healing powers might make “in body” regeneration of muscle tissue is a possibility.

Sang Jin Lee, assistant professor of Medicine at Wake Forest, and his colleagues implanted small bits of biomaterial scaffolds into the legs of rats and mice. When they embedded these scaffolds with proteins that mobilize muscle stem cells (like insulin-like growth factor-1 or IGF-1), the stem cells migrated from the muscles to the bioscaffolds and formed muscle tissue.

“Working to leverage the body’s own regenerative properties, we designed a muscle-specific scaffolding system that can actively participate in functional tissue regeneration,” said Lee. “This is a proof-of-concept study that we hope can one day be applied to human patients.”

If patients have large sections of muscle removed because of infections, tumors or accidents, muscle grafts from other parts of the body are typically used to restore at least some of the missing muscle. Several laboratories are trying the grow muscle in the laboratory from muscle biopsies that can be then transplanted back into the patient. Growing muscle on scaffolds fashioned from biomaterials have also proven successful.

Lee’s technique overcomes some of the short-comings of these aforementioned procedures. As Lee put it, “Our aim was to bypass the challenges of both of these techniques and to demonstrate the mobilization of muscle cells to a target-specific site for muscle regeneration.”

Most tissues in our bodies contain a resident stem cell population that serves to regenerate the tissue as needed. Lee and his colleagues wanted to determine if these resident stem cells could be coaxed to move from the tissue or origin, muscle in this case, and embeds themselves in an implanted scaffold.

In their first experiments, Lee and his team implanted scaffolds into the leg muscles of rats. After retrieving them several weeks later, it was clear that the muscle stem cell population (muscle satellite cells) not only migrated into the scaffold, but other stem cell populations had also taken up residence in the scaffolds. These scaffolds were also contained an interspersed network of blood vessels only 4 weeks aster transplantation.

In their next experiments, Lee and others laced the scaffolds with different cocktails of proteins to boost the stem cell recruitment properties of the implanted scaffolds. The protein that showed the most robust stem cell recruitment ability was IGF-1. In fact, IGF-1-laced scaffolds had four times the number of cells as plain scaffolds and increased formation of muscle fibers.

“The protein [IGF-1] effectively promoted cell recruitment and accelerated muscle regeneration,” said Lee.

For their next project, Lee would like to test the ability of his scaffolds to promote muscle regeneration in larger laboratory animals.

Pretreatment of Mesenchymal Stem Cells with Melatonin Improves Their Healing Properties in Animals with Strokes


The transplantation of mesenchymal stem cells or MSCs as they as affectionately known, does indeed benefit patients who have had a stroke. Unfortunately, the benefits of MSC transplantation if is limited by inability of these cells to survive after they are implanted into a low-oxygen environment. When a person suffers from a stroke, a blood vessel that feeds the brain has been blocked, and this blockage results in the death of particular cells in the brain. The affected areas of the brain, however, have been deprived of oxygen, and the transplantation of new cells into these areas can result in the prompt death of the implanted cells.

Fortunately, previous studies have revealed that pretreatment of the implanted cells with the hormone melatonin can increase the survival of MSCs that were implanted into kidneys that suffered oxygen deprivation. Therefore, could melatonin pretreatment also improve MSC survival in the case of strokes?

A new study by Guo-Yuan Yang and his colleagues at the Med-X Research Institute in Shanghai, China has examined the effects of melatonin pretreatment on the survival of MSCs that were implanted into the brains of laboratory animals that suffered a stroke.

In a nutshell, Yang and his colleagues showed that melatonin pretreatment greatly increased survival of cultured MSCs when these cells were subjected to low-oxygen conditions. Then when they went whole hog and transplanted their melatonin-pretreated MSCs into the brains of animals that had suffered a stroke, they once again observed that these cells survived at a substantially higher rate than their untreated counterparts. Melatonin-pretreated MSCs also further reduced bleeds into the brain (infarction) and improved the behavioral outcomes of the laboratory animals.

When Yang’s group examined the molecules secreted by the melatonin-treated MSCs, they discovered that the melatonin-pretreated MSCs made a lot more blood-vessel-promoting proteins (such as vascular endothelial growth factor or VEGF), and nerve cell-promoting molecules. Not surprisingly, the rats implanted with melatonin-pretreated MSCs shows significantly more new blood vessels formed, new neurons formed, and better looking brains in general.

Melatonin treatment increased the levels of two signaling molecules, p-ERK1/2, in MSCs. These particular signaling molecules are linked to higher survival rates. When Yang and his crew blocked melatonin signaling by treating cells with as drug called luzindole, these positive effects were reversed and when another drug called U0126, which prevents ERK from becoming phosphorylated was also applied to the cells, it completely reversed the protective effects of melatonin.

These results show that melatonin improves MSC survival and function. Furthermore, melatonin does this by activating the ERK1/2 signaling pathway. Therefore, mesenchymal cells pretreated by melatonin may represent a viable approach to enhance the beneficial effects of stem cell therapy for strokes, and maybe other conditions too? Well shall see. Stay tuned…..

Toxic stem cells to fight tumors


A Harvard team has developed special stem cells that secrete toxins that kill cancer cells, and cause no harm to healthy ones.

“Now, we have toxin-resistant stem cells that can make and release cancer-killing drugs,” Khalid Shah, a co-author of the study and the director of the Molecular Neurotherapy and Imaging Lab at Massachusetts General Hospital and Harvard Medical School, said in an official statement.

According to Shah, experiments in mice have proven very successful.

During the tests, the main part of the brain tumor was surgically removed, followed by the application of stem cells that were placed at the site of the tumor embedded in a biodegradable gel to kill the remaining cancerous cells.

Once within the cancer cell, the toxin disrupts its ability to synthesize proteins, killing it in a matter of days.

“After doing all of the molecular analysis and imaging to track the inhibition of protein synthesis within brain tumors, we do see the toxins kill the cancer cells,” he declared.

Shah said that the toxins that kill cancer have been used to treat a few types of blood cancers. However, these toxins were not effective dealing with solid tumors because these cancers are not as accessible and the toxins in the stem cells don’t have enough time to kill the cancer, since they only have a short half-life.

However, the new modified stem cells developed by Shah’s team change this limitation. “Now, we have toxin-resistant stem cells that can make and release cancer-killing drugs,” he said.

The study, published in the journal Stem Cells, possibly represents a breakthrough in cancer research, since it kills cancer cells while keeping remaining, healthy cells intact.

Scientists have applied for approval from the FDA to start the clinical trials of the method.

Experts praised the study as “the future” of cancer research.

“This is a clever study, which signals the beginning of the next wave of therapies. It shows you can attack solid tumors by putting minipharmacies inside the patient which deliver the toxic payload direct to the tumor,” Chris Mason, a professor of regenerative medicine at University College London, who was not participating in the study, told the BBC.

Human articular cartilage defects can be treated with nasal septum cells


A report from collaborating research teams from the University and the University Hospital of Basel specifies that cells isolated from the nasal septum cartilage can adapt to the environment the knee and repair articular cartilage defects. The ability of nasal cartilage cells to self-renew and adapt to the joint environment is associated with the expression of genes know as HOX genes. This research was published in the journal Science Translational Medicine in combination with reports of the first patients treated with their own nasal cartilage.

Lesions in articular or joint-specific cartilage is a degenerative that tends to occur in older people or younger athletes who engage in impact-heavy sports. Sometimes people who have experienced accidents can also suffer from cartilage lesions. Cartilage lesions present several challenges for orthopedic surgeons to repair. These surgeries are often complicated, and the recovery times are also long. However, Prof. Ivan Martin, professor of tissue engineering, and Prof. Marcel Jakob, Head of Traumatology, from the Department of Biomedicine at the University and the University Hospital of Basel have presented a new treatment option for cartilage lesions that includes the use of nasal cartilage cells to replace cartilage cells in joints.

When grown in cell culture, cartilage cells extracted from the nasal septum (also known as nasal chondrocytes) have a remarkable ability to generate new cartilage tissue after their growth in culture. In an ongoing clinical study, the Basal research group have taken small biopsies (6 millimeters in diameter) from the nasal septa of seven of 25 patients below the age of 55 years. After isolating the cartilage cells from these cartilage samples, they cultured these cells and expanded them and applied them to a three-dimensional scaffold in order to engineer a cartilage graft with a specific size (30 x 40 millimeters).

Martin and his colleagues used these very cartilage grafts to treat the cartilage lesions in human patients. After removing the damaged cartilage tissue from the knee of several patients, their knees were treated with the engineered, tailored tissue from their noses.

Two previous experiments demonstrated the potential efficacy of this procedure. First, a previous clinical study conducted in cooperation with plastic surgeons and the Basel group used the same method to successfully reconstruct nasal wings affected by tumors.

Secondly, a preclinical study with goats whose knees were implanted with nasal cartilage cells showed that these cells were not only compatible with the knee-joint, but also successfully reconstituted the joint cartilage. Lead author of this study, Karoliina Pelttari, and her colleagues were quite surprised that the implanted nasal cartilage cells, which originate from a completely different set of embryonic cell types than the knee-joint were compatible. Nasal septum cells develop from neuroectodermal cells, which also form the nervous system and their self-renewal capacity is attributed to their lack of expression of some homeobox (HOX) genes. However, these same HOX genes are expressed in articular cartilage cells that are formed by mesodermal cells in the embryo.

“The findings from the basic research and the preclinical studies on the properties of nasal cartilage cells and the resulting engineered transplants have opened up the possibility to investigate an innovative clinical treatment of cartilage damage,” says Prof. Ivan Martin about the results. Several studies have confirmed that human nasal cells maintain their capacity to grow and form new cartilage despite the age of the patient. This means that older people could also benefit from this new method, as could patients with large articular cartilage defects.

The primary target of the ongoing clinical study at the University Hospital of Basel is to confirm the safety, efficacy and feasibility of nasal cartilage grafts transplanted into joints, the clinical effectiveness of this procedure, from the data presently in hand, is highly promising.

Patient’s Own Stem Cells Treat Rare Neurological Disorder


Stiff-Person syndrome is a rare neurological disease that, for all intents and purposes, looks like an autoimmune disease. It is characterized by muscular rigidity that tends to come and go. This rigidity occurs in the muscles of the trunks and limbs. Patients with Stiff-Person syndrome also have an enhanced sensitivity to stimuli such as noise, touch, and emotional distress, and various stimuli may cause the patient to experience painful muscle spasms that cause abnormal postures and stiffening. Stiff-Person syndrome or SPS is more common in women than in men and SPS patients often suffer from other autoimmune conditions in addition to SPS (for example, pernicious anemia, diabetes, vitiligo, and thyroiditis). Unfortunately, the precise cause of SPS is not known, but again, it looks like an autoimmune condition.

A research team at Ottawa Hospital Research Institute has made a breakthrough in the successful treatment of SPS using bone marrow stem cell transplants. The medical director at the Ottawa Hospital Research Institute, Dr. Harold L. Atkins, who is also a physician in the Blood and Bone Marrow Transplant Program at The Ottawa Hospital and an associate professor at the University of Ottawa has used bone marrow transplants to two female SPS patients into remission.

SPS can leave patients bedridden and in severe pain, but thanks to Atkins and his team, the progression of the disease in these women has ceased, allowing both women to regain their previous function and leaving them well enough to return to work and normal everyday activities.

Adkins and his group published this case study in JAMA Neurology, which is produced by the Journal of the American Medical Association. This is the first documented report that taking stem cells from a person’s own body can produce long-lasting remission of stiff person syndrome.

“We approach these cases very carefully and are always aware that there have just been a few patients treated and followed for a short time,” says Dr. Atkins. Atkins and his extracted bone marrow stem cells from each woman, and then used chemotherapy to eliminate their immune systems. Once their immune system were reliably eliminated, both women had their own stem cells returned to their bodies in order to reconstitute their immune systems. This procedure essentially gives the immune system a “do-over.”.

“By changing the immune system, one hopes to put the stiff person syndrome into remission,” adds Dr. Atkins. “Seeing these two patients return to their normal lives is really every physicians dream.”

This very procedure, which is known as an “autologous stem cell transfer” or ASCT has been used to successfully treat people who suffer from autoimmune diseases such as multiple sclerosis, scleroderma, and systemic lupus erythematosis. Atkins and his team used high-doses of chemotherapy and antibodies that specifically bind lymphocytes to rid the women’s bodies of their rogue immune cells before their immune systems were regenerated using their own stem cells. Adkins an his colleagues viewed this as a viable treatment option based strategies that had been used to treat other autoimmune diseases.

Patient 1 was diagnosed with stiff person syndrome in 2005 at age 48 after experiencing leg stiffness and several falls. After her treatment, her symptoms disappeared and she was fully mobile again six months after receiving the stem cell transplant procedure in 2009.

Patient 2 was diagnosed with stiff person syndrome in 2008 at age 30. She had stopped working and driving, and had moved back in with her parents before her stem cell transplant in 2011. Also, she has been able to return to her work and previous activities, and has not had any stiff person syndrome symptoms in more than a year.

“The results achieved by Dr. Atkins and his team through this innovative treatment show how research at The Ottawa Hospital can lead to life-changing and, even life-saving care,” says Dr. Duncan Stewart, Chief Executive Officer and Scientific Director of the Ottawa Hospital Research Institute. “Translating research into better care for patients is what we’re all about at the research institute.”

A More Efficient Way to Make Induced Pluripotent Stam cells


Mark Stadtfeld and his colleagues at the NYU Longone Medical Center has devised a new method for making induced pluripotent stem cells that greatly increases efficiency at which these cells are made.

Induced pluripotent stem cells or iPSCs are made from mature, adult cells by mean of a combination of genetic engineering and cell culture techniques. In short, the expression of four genes is forced in adult cells; Oct4, Sox2, Klf4, and c-Myc or OSKM. The proteins encoded by these four genes cooperatively work to drive a fraction of the cells into an immature state that resembles that of embryonic stem cells. These cells are them grown in cell culture systems that select for those cells that can grow continuously and form colonies of cells derived from progenitor cells. These cell colonies are them repeated isolated a re-cultured until an iPSC line has been established.

Unfortunately, this process is rather inefficient and tedious, since less than one percent or so of the reprogrammed cells actually undergo successful reprogramming. Additionally, it can take several weeks to properly establish an iPSC line. Thus, stem cell scientists have been looking at several different ways to boost the efficiency of this process.

Stadtfeld and his coworkers tried to add compounds to the cultured cells to determine if the culture conditions could actually augment the efficiency of the reprogramming process. “We especially wanted to know if these compounds could be combined to obtain stem cells at high-efficiency,” said Stadtfeld.

The compounds to which Stadtfeld was referring were two cell signaling proteins called Wnt and TFG-beta. Both of these compounds regulate a host of cell growth processes. Stadtfeld wanted to try regulating both of these pathways at the same time, in addition to providing cells with ascorbic acid, which is also known as vitamin C. Even vitamin C is more popularly known as an antioxidant, vitamin C also can remodel chromatin (that tight structure into which cells package their DNA).

When mouse skin fibroblasts were treated with OSKM and a compound that activates Wnt signaling, the efficiency of iPSC derivation increased slightly. The same thing was observed if fibroblasts were treated with OSKM and a compound that inhibits TGF-beta signaling or vitamin C. However, when all three of these compounds were combined, OSKM-engineered fibroblasts were reprogrammed at an efficiency of close to 80 percent. When different cell types were used as the starting cell, such as blood progenitor cells, the efficiency jumped to close to 100 percent; a result that was also observed if liver progenitor cells were used as the starting cell.

Stadtfeld is confident that these dramatic increases in iPSC derivation should improve future studies with iPSCs, since his protocol should make iPSC derivation more predictable. “It’s just a lot easier this way to study the mechanisms that govern reprogramming, as well as detect any undesired features that might develop in iPSCs,” he said.

Vitamin C and the two compounds used to manipulate the Wnt and TGF-β pathways have been widely used in research and have few unknown or hazardous effects. However, OKSM has in some cases caused undesired features in iPSCs, such as increased mutation rates. Stadtfeld believes that by making iPSC induction more rapid and efficient, his new technique might also make the resulting stem cells safer. “Conceivably it reduces the risk of abnormalities by smoothening out the reprogramming process,” Dr. Stadtfeld says. “That’s one of the issues we’re following up.”