Umbilical Cord Stem Cells Preserve Heart Function After a Heart Attack in Mice


A consortium of Portuguese scientists have conducted an extensive examination of the effects of mesenchymal stromal cells from umbilical cord on the heart of mice that have suffered a massive heart attack. Even more remarkable is that these workers used a proprietary technique to harvest, process, and prepare the umbilical cord stem cells in the hopes that this technique would give rise to a commercial product that will be tested in human clinical trials,

Human umbilical cord tissue-derived Mesenchymal Stromal Cells (MSCs) were obtained by means of a proprietary technology that was developed by a biomedical company called ECBio. Their product,, UCX®, consists of clean, high-quality, umbilical cord stem cells that are collected under Good Manufacturing Practices. The use of Good Manufacturing Practice means that UCX is potentially a clinical-grade product. Thus, this paper represents a preclinical evaluation of UCX.

This experiments in this paper used standard methods to give mice heart attacks that were later received injections of UCX into their heart muscle. The same UCX cells were used in experiments with cultured cells to determine their effects under more controlled conditions.

The mice that received the UCX injections into their heart muscles after suffering from a large heart attack showed preservation of heart function. Also, measurements of the numbers of dead cells in the heart muscle of heart-sick mice that did and did not receive injections of umbilical cord cells into their hearts showed that the umbilical cord stem cells preserved heart muscle cells and prevented them from dying. Additionally, the implanted umbilical cord MSCs induced the growth and formation of many small blood vessels in the infarcted area of the heart. This prevented the heart from undergoing remodeling (enlargement), and preserved heart structure and function.

When subjected to a battery of tests on cultured cells, UCX activated cardiac stem cells, which are the resident stem cell population in the heart. Implanted UCX cells activated the proliferation of cardiac stem cells and their differentiation into heart muscle cells. There was no evidence that umbilical cord MSCs differentiated into heart muscle cells and engrafted into the heart. Rather UCX seems to help the heart by means of paracrine mechanisms, which simply means that they secrete healing molecules in the heart and help the heart heal itself.

In conclusion, Diana Santos Nascimento, the lead author of this work, and her colleagues state that, “the method of UCX® extraction and subsequent processing has been recently adapted to advanced therapy medicinal product (ATMP) standards, as defined by the guideline on the minimum quality data for certification of ATMP. Given that our work constitutes a proof-of-principle for the cardioprotective effects UCX® exert in the context of MI, a future clinical usage of this off-the-shelf cellular product can be envisaged.”

Preclinical trials with larger animals should come next, and after that, hopefully, the first human clinical trials will begin.

Artificial Skin Created Using Umbilical Cord Stem Cells


Major burn patients usually must wait weeks for artificial skin to be grown in the laboratory to replace their damaged skin, buy a Spanish laboratory has developed new protocols and techniques that accelerate the growth of artificial skin from umbilical cord stem cells. Such laboratory-grown skin can be frozen and stored in tissue banks and used when needed.

Growing skin in the laboratory requires the acquisition of keratinocytes, those cells that compose the skin and the mucosal covering inside our mouths.  Keratinocytes can be cultured in the laboratory, but they have a long cell cycle, which means that they take a really long time to divide.  Consequently, cell cultures of keratinocytes tend to take a very long time to grow.

Keratinocytes in culture
Keratinocytes in culture

As they grow, the keratinocytes respond to connective tissue underneath them to receive the cues that tell them how to connect with each other and form either skin or oral mucosa.  In patients with severe burns, however, the underlying connective tissue is also often damaged.  Therefore, finding a way to not only accelerate the growth of cultured keratinocytes, but also to provide the underlying structure that directs the cells to form a proper epithelium is essential.

Remember that severe burn patients are living on borrowed time.  Without a proper skin covering, water loss is severe and dehydration is a genuine threat.  Also, infection is another looming threat.  Therefore, the treatment of a burn patient is a race against time.

Because umbilical cord stem cells grow quickly and effectively in culture, they might be able to differentiate into keratinocytes and form the structures associated with oral mucosa and skin.

University of Granada researchers used a new type of epithelial covering to grow their artificial skin in addition to a biomaterial made of fibrin (the stiff, cable-like protein that forms clots) and agarose to provide the underlying connective tissue. In case you might need a refresher, an epithelium refers to a layer of cells that have distinct connects with each other and form a discrete layer. Epithelia can form single or multiple layers and can be composed of long, skinny cells, short, flat cells, or boxy cells.  An epithelium is a membrane-like tissue composed of one or more layers of cells separated by very little intervening substances.  Epithelia cover most internal and external surfaces of the body and its organs.

Previous work from this same research group showed that stem cells from Wharton’s jelly (connective tissue within the umbilical cord), could be converted into epithelial cells. This current study confirms and extends this previous work and applies it to growing skin, and oral mucosa.

“Creating this new type of skin suing stem cells, which can be stored in tissue banks, mains that it can be used instantly when injuries are caused, and which would bring the application of artificial skin forward many weeks,” said Antonio Campos, professor of histology and one of the authors of this study.

By growing the Wharton’s jelly stem cells on their engineered matrix in a three-dimensional culture system, Campos and his colleagues saw that the stem cells stratified (formed layers), and expressed a bunch of genes that are peculiar to skin and other types of epithelia that cover surfaces (e.g., cytokeratins 1, 4, 8, and 13; plakoglobin, filaggrin, and involucrin).  When examined with an electron microscope, the cells had truly formed the kinds of tight connections and junctions that are so common to skin epithelia.

Electron microscopy analysis of controls and three-dimensional bioactive models of H-hOM and H-hS. SEM images (top) corresponding to N-hOM and N-hS controls showed a tight superficial layer of flat polygonal cells with desquamation signs in which cells were covering the entire surface, whereas samples kept in vitro for 2 weeks showed immature differentiation patterns, and samples implanted in vivo for 40 days tended to resemble the structure of the native control tissues, with flattened cells and evident signs of desquamation. Scale bars = 50 μm. TEM samples (bottom) were analyzed after 40 days of in vivo implantation and demonstrated that in vivo-implanted tissues were mature and well-differentiated, with numerous intercellular junctions, abundant cell organelles, and a collagen-rich stroma. Scale bars = 1 μm. Abbreviations: H-hOM, heterotypical human oral mucosa; H-hS, heterotypical human skin; N-hOM, native human oral mucosa; N-hS, native human skin; SEM, scanning electron microscopy; TEM, transmission electron microscopy.
Electron microscopy analysis of controls and three-dimensional bioactive models of H-hOM and H-hS. SEM images (top) corresponding to N-hOM and N-hS controls showed a tight superficial layer of flat polygonal cells with desquamation signs in which cells were covering the entire surface, whereas samples kept in vitro for 2 weeks showed immature differentiation patterns, and samples implanted in vivo for 40 days tended to resemble the structure of the native control tissues, with flattened cells and evident signs of desquamation. Scale bars = 50 μm. TEM samples (bottom) were analyzed after 40 days of in vivo implantation and demonstrated that in vivo-implanted tissues were mature and well-differentiated, with numerous intercellular junctions, abundant cell organelles, and a collagen-rich stroma. Scale bars = 1 μm. Abbreviations: H-hOM, heterotypical human oral mucosa; H-hS, heterotypical human skin; N-hOM, native human oral mucosa; N-hS, native human skin; SEM, scanning electron microscopy; TEM, transmission electron microscopy.

The authors conclude the article with this statement: “All these findings support the idea that HWJSCs could be useful for the development of human skin and oral mucosa tissues for clinical use in patients with large skin and oral mucosa injuries.”  Think of it folks – new skin for burn patients, quickly, safely and ethically.

Now back to reality – this is exciting, but it is a a pre-clinical study.  Larger animals studies must show the efficacy and safety of this protocol before human trials can be considered, but you must admit that it looks exciting; and without killing any embryos.

See I. Garzón, et al., Stem Cells Trans MedAugust 2013 vol. 2 no. 8625-632.

Artificial Bones From Umbilical Cord Stem Cells


I am back from vacation. We visited some colleges in Indiana for my daughter who will be a senior this year. She really liked Taylor University and Anderson University. We’ll see if the tuition exchange works out.

Now to blogging.

Scientists from Granada, Spain have patented a hew biomaterial that consists of activated carbon cloth that just happens to be able to support the growth of cells that have the ability to regenerate bone. These results came from experiments that were conducted outside any living animals, but they hope to confirm these results in a living animal in the near future.

This new biomaterial facilitates the growth of bone-making cells derived from umbilical cord stem cells. This activated carbon cloth acts as a scaffold for cells that differentiate into “osteoblasts,” which are bone-building cells. This activated carbon cloth gives the osteoblasts a proper surface upon which to promote the growth of new bone.

Bone loss as a result of cancer, trauma, or degenerative bone diseases requires replacement bone to heal to damaged bone. Making new bone in the laboratory that can be transplanted is an optimal strategy for treating these patients.

Even though this laboratory-made bone was not used in living laboratory animals to date, the laboratory results look quite impressive. In the future, such techniques could help manufacture medicines or other sources of material to repair bone or lost cartilage. Once such artificial bone has been made in the laboratory, the Spanish team hopes to transplant it into rats or rabbits to determine if it can regenerate bone in such creatures.

Presently, no materials exist to replace lost bone. The method used to make bone by the research team from Granada uses a three-dimensional support that facilitates the production of those cell types that regenerate bone without the need for additional growth factors.

The growth of these umbilical cord stem cells on activated carbon cloth produced a product that could produce organic bone, but also mineralize the organic bone matrix. This patent could have numerous clinical applications in regenerative medicine and the Granada group hopes to obtain funding to continue this work and achieve their ultimate objective: to regenerate bones by implanting biomaterial in patients with bone diseases.

Umbilical Cord Blood Stem Cells Revive Child From Persistent Vegetative State


Physicians from Ruhr-Universitaet-Bochum (RUB) have successfully treated cerebral palsy in a 2.5-year old boy with his own cord blood.

“Our findings, along with those from a Korean study, dispel the long-held doubts about the effectiveness of the new therapy,” says Dr. Arne Jensen of the Campus Clinic gynaecology. Jensen collaborated with his colleague Prof. Dr. Eckard Hamelmann of the Department of Pediatrics at the Catholic Hospital Bochum (University Clinic of the RUB). This case study was published in the journal Case Reports in Transplantation.

At the end of November 2008, a young child’s heart stopped (cardiac arrest), and his brain suffered oxygen deprivation, and, consequently, severe brain damage. He was in a persistent vegetative state, and his body was completely paralyzed. This condition, infantile cerebral palsy, until now, has no recognized treatment. Typically, the prognosis of children with infantile cerebral palsy is rather grim, since the chances of survival miniscule and months after suffering severe brain damage, the surviving children usually only exhibit minimal signs of consciousness. According to the physicians at RUB, “The prognosis for the little patient was threatening if not hopeless.”

However, this child’s persistent parents scoured the literature for alternative therapies to infantile cerebral palsy. Arne Jensen explains. “They contacted us and asked about the possibilities of using their son’s cord blood, frozen at his birth.”

Nine weeks after suffering brain damage, on 27 January 2009, Jensen and his colleagues administered the child’s prepared cord blood intravenously. They studied the child’s progressive recovery at 2, 5, 12, 24, 30, and 40 months after treatment.

After the cord blood therapy, the patient, however, recovered quickly. Within two months, the child’s spasms decreased significantly. He was able to see, sit, smile, and to speak simple words again. Forty months after treatment, the child was able to eat independently, walk with assistance, and form four-word sentences. “Of course, on the basis of these results, we cannot clearly say what the cause of the recovery is,” Jensen says. “It is, however, very difficult to explain these remarkable effects by purely symptomatic treatment during active rehabilitation.”

Just listen to the description of the child’s recovery from this paper:

After two years, there was independent eating and speech competence of eight words (pronunciation slurred, mimicking prosody) with broad understanding. The patient moved from a prone to a free sitting position and crawled without cross-pattern, but using the arms. Independent passive standing, walking with support, and independent locomotion in a gait trainer was possible (video S5). He played imaginative games, and recognized colours, animals, and objects, assigning them correctly. Fine motor control improved to such an extent that he managed to steer a remote control car (video S6). At 30 months, he formed two-word-sentences using 80 words.

After 40 months, there was further improvement in both receptive and expressive speech competence (four-word-sentences, 200 words), walking (Crocodile Retrowalker), crawling with cross-pattern, and getting into vertical position.

And this is from a child who was a in a persistent vegetative state, who could neither speak, nor eat on his own, nor talk.

In animal studies, scientists have examined the therapeutic potential of cord blood. In a previous study with rats, RUB researchers revealed that cord blood cells migrate to the damaged area of the brain in large numbers within 24 hours of administration.  Umbilical cord stem cells are also known to secrete gobs of neurotropic molecules that stimulate neuron growth and differentiation, promote neuron survival, quell inflammation, staunch star formation in the brain (gliosis), and stimulate the growth and formation of blood vessels.

In March 2013, in a controlled study of one hundred children, Korean doctors reported for the first time that they had successfully treated cerebral palsy with someone else’s cord blood.

These results show that cord blood has tremendous therapeutic potential for pediatric neurological conditions.  This remarkable recovery is seemingly miraculous.  Certainly this merits more work and excitement.

Hair-like Structures Formed by Umbilical Cord Stem Cells


Michael Detamore‘s laboratory at the University of Kansas Medical Center has used mesenchymal stem cells from connective tissue in human umbilical cord tissue to form structures that have some although not all features of human hair.

Human umbilical cord contains a unique connective tissue called “Wharton’s jelly.” Wharton’s jelly expands after birth to staunch bleeding from the umbilical veins and arteries. Within Wharton’s jelly is a mesenchymal stem cell population called Wharton’s Jelly Mesenchymal Stromal Cells or WJMSCs. Experiments in a variety of labs have shown that WJMSCs have differentiate into bone, cartilage, muscle, and neural cells. Recently, several labs have used three-dimensional culture systems to get WJMSCs to differentiate into tubular and endometrial cells.

Detamore and his co-workers used this three-dimensional culture system to differentiate WJMSCs into layered structures that expressed some of the genes associated with hair follicles and also looked like follicles. The cells formed small spheres of cells known as “spheroids.” Detamore and his colleagues showed that these spheroids can form bone, but during done differentiation, Detamore and the other workers in his lab noticed that the spheroids form protrusions that looked like hairs. See for yourself below.

Patterns of hair-like structures in DWJM. Hair-like structures were observed growing out of DWJM 2 weeks following WJMSC seeding and osteogenic induction. In phase-contrast microscopy pictures (A, B) taken during culture, hair-like structures were observed to grow under the outer layer of the DWJM and in some cases either successfully protruded through the outer layer (A) or just caused a protrusion of the outer layer of DWJM (B). Also, the hair-like structures were either straight (A, C–E) or coiled (B). The green arrows in picture (A) pointed to the outer layer covering DWJM. The hair-like structure caused the outer layer covering DWJM to appear lifted up. In this DWJM, 2 hair-like structures protruded through DWJM (red arrows) seen in (A, C–E). A second hair-like structure (red arrows) protruded through the outer layer of DWJM; however, this hair-like structure was surrounded by tissue material (E). In (F), multiple areas of protrusions noted (black arrows). In (C–E) pictures, material was visualized using Nikon SMX1500 dissecting microscope and pictures were taken using Optem DC50NN camera. Scale bars represent 100 μm.
Patterns of hair-like structures in DWJM. Hair-like structures were observed growing out of DWJM 2 weeks following WJMSC seeding and osteogenic induction. In phase-contrast microscopy pictures (A, B) taken during culture, hair-like structures were observed to grow under the outer layer of the DWJM and in some cases either successfully protruded through the outer layer (A) or just caused a protrusion of the outer layer of DWJM (B). Also, the hair-like structures were either straight (A, C–E) or coiled (B). The green arrows in picture (A) pointed to the outer layer covering DWJM. The hair-like structure caused the outer layer covering DWJM to appear lifted up. In this DWJM, 2 hair-like structures protruded through DWJM (red arrows) seen in (A, C–E). A second hair-like structure (red arrows) protruded through the outer layer of DWJM; however, this hair-like structure was surrounded by tissue material (E). In (F), multiple areas of protrusions noted (black arrows). In (C–E) pictures, material was visualized using Nikon SMX1500 dissecting microscope and pictures were taken using Optem DC50NN camera. Scale bars represent 100 μm.

The other side of the spheroid did form bone, but the hair-like structures did eventually form bone. There are specific genes that are expressed in hair follicles, and these can be used to determine if the projections are actually hair follicles. One of these genes, cytokeratin 19 or CK19, was expressed at pretty high levels in the hair follicles. Another hair-specific gene, CK15 was also expressed in the hair-like structures.

Are these real hair follicles? Probably not, but they seem to be on their way to making hair follicles. Furthermore, the production of these hair-like structures was rather easy. If WJMSCs could be used to make hair, then they might be useful for cosmetic procedures that replace lost hair follicles as a result of baldness.

Umbilical Cord Stem Cells Outperform Bone Marrow Stem Cell in Heart Repair


A study from the laboratory of Armand Keating at the University of Toronto and Princess Margaret Hospital has compared the ability of umbilical cord stem cells and bone marrow stem cells to repair the hearts of laboratory animals after a heart attack. The umbilical cord stem cells showed a clear superiority to bone marrow stem cells when it came to repairing heart muscle.

Keating used human umbilical cord perivascular cells (HUCPVCs) for his experiment, and these cells are widely regarded as a form of umbilical cord mesenchymal stem cell that surround the umbilical cord blood vessels.

Transplantation of cells from either bone marrow or umbilical cord into the heart soon after a heart attack improved the function and structure of the heart. However, functional measurements showed that the HUCPVCs were twice as effective as bone marrow stem cells at repairing the heart muscle.

Keating added: “We are hoping that this translates into fewer people developing complications of heart failure because their muscle function after a heart attack is better.”

In addition to further pre-clinical tests, Keating and his research team hope to initiate clinical trials with human patients within 12-18 months. Keating is also interested in testing the ability of umbilical cord stem cells to heal the hearts of those cancer patients who have experienced heart damage as a result of chemotherapy. In such patients, chemotherapy rids their bodies of cancer, but the cure is worse than the cancer, since the drugs also leave the patients with a severely damaged heart. Such stem cell transplantations could potentially strengthen the hearts of these patients, and give them a new lease on life. My own mother died from congestive heart failure as a result of an experimental arsenic treatment that killed her heart muscle. My mother suffered from chronic myelogenous disease and the arsenic was meant to kill off all the rogue cells in her bone marrow, but instead it killed her heart. If such a stem treatment were available then, my mother might still be with me.

There are over 250 clinical trials with mesenchymal stem cells to date to treat conditions ranging from Crohn’s disease to neurological conditions.  Also, a recent meta-analysis has established the safety of mesenchymal stem cell treatments for several different conditions (see Lalu MM, McIntyre L, Pugliese C, Fergusson D, Winston BW, et al. (2012) Safety of Cell Therapy with Mesenchymal Stromal Cells (SafeCell): A Systematic Review and Meta-Analysis of Clinical Trials. PLoS ONE 7(10): e47559. doi:10.1371/journal.pone.0047559).

Stem Cell Therapy for Inflammatory Bowel Disease in the Works


Stem cells from umbilical cord blood have the ability to migrate to the intestine and integrate into the tissues. This integration allows umbilical stem cells to contribute to the cell population of the gastrointestinal tract. This biological property of umbilical cord stem cells might make them ideal treatments for inflammatory bowel disease (IBD).

One million Americans have IBDs such as Crohn’s disease or ulcerative colitis. Crohn’s disease can affect the small and large intestine, whereas the ulcerative colitis is usually restricted to the colon (large intestine). Also Crohn’s disease displays patchy lesions whereas ulcerative colitis consists of continuous stretches of inflammation. These disease are characterized by frequent diarrhea and abdominal pain. Patients who suffer from ulcerative colitis also tend to have bloody stools, and if left untreated, the blood loss can be extensive. Ulcerative colitis only affects the upper layer of the large intestine, whereas Crohn’s disease can affect multiple layers of the intestine.

There are no cures for IBDs, but there are drug treatments. In the case of ulcerative colitis, the drug prednisone is used to calm down fulminant outbreaks and then mesalamine (5-aminosalicylic acid) or sulfasalazine are used to maintain the disease in a calm or quiescent state. Mesalamine is present in an oral form marketed as Asacol or Pentasa. The difference between Asacol and Pentasa is in the outer chemical coating, since Pentasa packages its drug in coated microgranules, which enables a prolonged release of the active substance throughout the intestinal tract, from duodenum to the rectum. Therefore Pentasa is more useful for Crohn’s patients. Asacol is a delayed release enteric-coated tablets that releases the active ingredient only in the colon. Mesalamine is also available in an enema form (Rowasa)

If these drugs do not work, biologic treatments such as Infliximab (Remicade), adalimumab (Humira) and Golimumab (Simponi) are commonly used to treat patients with Ulcerative Colitis, but these drugs suppress the immune system and can raise the risk of severe illness. Corticosteroids are also used, but long-term use of these drugs also causes severe side effects.

Thus, if the drugs do not work, the treatment can be as bad as the disease itself. Certainly a treatment that regenerates the bowel is preferable, and a stem cell treatment seems to fit the bill.

In an article in the journal Hepatology, the senior author, Graca Almeida-Porada, a professor at Wake Forest Baptist Medical Center’s Institute for Regenerative Medicine, and her colleagues argue that a special stem cell population known as endothelial colony-forming cells, found in umbilical cord blood and bone marrow and circulating blood, can play a definite role in the treatment of IBDs.

Almeida-Porada said, “These cells are involved in the formation of blood vessels and may prove to be a tool for improving the vessel abnormalities found in IBD.”

In 1997, scientists discovered that these endothelial colony-forming stem cells contribute to the formation of blood vessels in embryos, and adults. This study initiated investigations of the capacity of endothelial colony-forming cells as potential therapeutic agents. Clinical studies have shown that endothelial colony-forming cells can improve reduced blood flow to limbs and can also treat heart disease.

Unfortunately, few studies have examined the ability of endothelial colony-forming cells to home to different organs and integrate into their circulatory systems. Thus, Almeida-Porada wanted to examine the ability of endothelial colony-forming cells to integrate into the intestine. Also, since abnormal blood vessels are a hallmark of IBDs, they might be a potential treatment for IBDs.

In this experiment, fetal sheep at 59-65 days gestation were injected with human endothelial colony-forming cells (EPCs). At 11 weeks gestation, the fetal sheep were examined to determine if the human cells had integrated into the fetal sheep tissue. Researchers found that the infused cells had migrated into the intestine and had made significant contributions to the cell population of the bowel.

According to Almeida-Porada: “The study shows that the cells can migrate to and survive in a healthy intestine and have the potential to support vascular health. Our next step will be to determine whether cells can survive in the ‘war’ environment of an inflamed intestine.”

Interestingly, Almeida-Porada’s team found that endothelial colony-forming cells also colonized the liver of the fetal sheep. Although smaller numbers of cells reached the liver as opposed to the intestine, new strategies might enhance the therapeutic potential for these cells with respect to the liver.