Thursday, September 20, 2012

Your Body's Stem Cell Bank Account

By Neil Riordan, PhD

Everybody is born with a certain amount of stem cells, specifically, "adult stem cells." This number may be thought of as a type of "bank account", from which each person may make "withdrawals" throughout his or her lifetime, as needed.

However, not all bank accounts are created equal. To continue with the analogy, some people are born rich, while others are born poor. Most people, however, may be thought of as "middle class." This fact helps to explain why some people are able to enjoy health and longevity despite very unhealthy lifestyles, while other people may enjoy neither robust health nor longevity despite healthy lifestyles. In other words, some people are able to "spend" their stem cells more extravagantly than others, simply because they have more to spend. Most of us, however, fall somewhere in the middle, where both the length and the quality of our lives may be influenced to some degree by our choice of lifestyle. Environmental factors also play a key role in determining how rapidly one's "bank account" of (adult) stem cells is depleted. However, even under ideal circumstances, stem cells continually diminish with age.

Our stem cells exist in every part of the body to repair damage, such as from broken bones, a paper cut, radiological or chemical exposure, etc., all of which require stem cells for healing. You may draw on your bank account at any time, whenever you need to do so, until you run out of stem cells. As your "bank account" of stem cells approaches zero, physiological healing will become increasingly difficult, until finally it ceases altogether.

Utilizing stem cells is like going to the ATM machine. Depending upon how you live your life, and whether you were born with a large or a small bank account of stem cells, you either may or may not be able to withdraw from your account. If you were fortunate enough to be born with a large amount of stem cells, then you might possibly be able to smoke and drink and eat unhealthy food and never exercise, and still live to a ripe old age, because regardless of how many stem cells you expend, there are still more to be spent.

On the other hand, if you were born at the opposite end of the spectrum, with a small amount of stem cells, then an unhealthy lifestyle will have a more immediate and detrimental impact upon the quality and length of your life. This is known as Nature's Law of Conservancy: the less stem cells that exist in your "account", the more stingy the ATM machine becomes in distributing the contents of that account. Most people, however, are born into the stem cell "middle class", which is to say that lifestyle choices can often make a noticeable difference in determining health and longevity.

When someone has a large number of stem cells in the bank, the ATM works very quickly and efficiently. But when the bank account is almost empty, which ordinarily happens in the latter years of life, the ATM machine does not distribute the stem cells as readily. In biological terms, this is because the division rate of the cells has slowed considerably. Simply changing the doubling time of stem cells from 24 hours to 72 hours can make a 90-day difference in the amount of time required to reach a "critical mass" of cells that are required to heal a wound. Michael Andreeff, M.D., Ph.D., a professor in the Departments of Blood and Marrow Transplantation and Leukemia Cancer at M.D. Anderson, has described cancer as a "never healing wound"; in some cases, the doubling time of the stem cells may never be fast enough to "catch up" and heal the wound. This is why the incidence of cancer increases with age. After childhood cancers (which arise due to genetic factors or to overwhelming environmental exposure), the incidence of cancer drops significantly until around the age of 40, when it starts to increase, shooting up dramatically around the age of 50, and then falling again around the age of 65.

When someone has fully depleted his or her stem cell reserve, the only possible way to get more stem cells is from an alternate source. This is where stem cell therapy comes into play.


Salamanders are a supreme example of stem cell billionaires. Salamanders have a seemingly unlimited supply of stem cells, as their "bank accounts" are virtually incapable of being depleted. If you were to look at the blood of a salamander under a microscope, you would see that all of the red blood cells (RBCs) are nucleated. In other words, all RBCs of a salamander contain nuclei, unlike human RBCs, which are not nucleated. This is why salamanders can regenerate entire limbs and humans cannot: every RBC in a salamander is a functional stem cell, floating around everywhere throughout the salamander's body. In humans, by contrast, our RBCs cannot replicate themselves once they have migrated out of our bone marrow. As long as the salamander is still alive, its ATM machine is fast and generous in distributing as many stem cells as may be needed for any task.

Although humans are not salamanders, the ability to regenerate entire limbs nevertheless offers a powerful example of the possible applications and implications of stem cells. Practically as well as theoretically, an enormous, mostly untapped, potential exists in the field of stem cell therapy.

Inflammatory Bowel Disease Treatable With Stem Cells?

Researchers at Wake Forest Baptist Medical Center's Institute for Regenerative Medicine may have discovered the key to treating inflammatory bowel disease (IBD).

Dr. Graca Almeida-Porada and her team of scientists found a specific stem cell population in cord blood that migrates to the intestine and proliferates there.
Fetal sheep were injected with the stem cells and their intestines were analyzed 11 weeks later.

"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," said Dr. Almeida-Porada.

Intestinal swelling, inflammation and ulcers typically cause abdominal pain and diarrhea in IBD patients. Reducing inflammation is a key to treatment but currently approved drugs are not very effective.

"This study shows that the cells can migrate to and survive in a healthy intestine and have the potential to support vascular health," said Almeida-Porada. "Our next step will be to determine whether the cells can survive in the 'war' environment of an inflamed intestine."

Friday, September 14, 2012

Stem cell treatment in Panama benefits autistic Glenburn youth


By Dale McGarrigle, Of The Weekly Staff
Bangor Daily News
Posted Sept. 14, 2012, at 12:17 p.m.

GLENBURN — Now Kenny can read.

Kenny Kelley can now also do many things that other 11-year-olds take for granted. According to his mother, Marty Kelley, that’s because injections of adult stem cells, taken from umbilical cord blood, have helped Kenny to shake off the shackles of autism, with which he was first diagnosed at age 2.

“The results from stem cells can be seen everyday in his amazing thoughts and vast imagination!!,” Marty Kelley wrote in her blog, http://www.kensjourneytorecovery.blogspot.com/. “How lucky we are for such a miracle treatment!”

Autism is a brain disorder found in children that interferes with their ability to communicate and relate to other people. Autism affects 1 in 88 children and 1 in 54 boys. What causes autism has not been established.

Stem cells are the body’s internal repair system and can fix and replace damaged tissue. These unspecialized cells are a blank slate, capable of transforming into muscle cells, blood cells, and brain cells. Stem cells can also renew themselves by dividing and giving rise to more stem cells.

Stem cells taken from umbilical cord blood, such as Kenny received, are the least likely to be rejected.

The stem-cell treatment is the latest effort by Marty and her husband, Donald, to find ways to improve Kenny’s life. The Kelleys also have two other children: Philip, 13, and Caroline-Grace, 6.

First was in-home treatment in a mild hyperbaric oxygen chamber, three hours a day equaling 800 hours over the course of two years, beginning when Kenny was 5 ½ to 6 years old. This was coupled with a Specific Carbohydrate Diet, which restricts the use of complex carbohydrates and eliminates refined sugar and all grains and starch from the diet.

“We saw results right away with the chamber,” Marty recalled in a recent interview. “He made slow gains, such as tracing the alphabet.”

Then the Kelleys discovered on the Internet the story of Matthew Faiella, a New York boy who has been making great strides after stem-cell treatment in Panama for his autism. They decided to follow suit.

Why take this path, when there has been little scientific research into the use of stem cells to treat autism?

“We were willing to do it as long as it’s safe, and I’ve researched this,” Marty said. “Stem cells are very natural. I’m not a scientist, but I care much more than any scientist would, and I would never do anything to hurt my baby.”

When Kenny went for his first stem-cell treatment in July 2009, at the Stem Cell Institute in Costa Rica, Marty assessed the condition of her then 8-year-old son in her blog http://www.kensjourneytorecovery.blogspot.com:

• Behavior: Screaming, aggressive, giggles/silly/inappropriate with his brother or new people, running around, destructive, uncooperative while being dressed, hitting, not potty trained (still wearing diapers).

• Speech: Vocabulary of a 4-year-old. He can talk, but it is difficult for strangers to understand him. Answers some questions, but he does not understand or like why, when, or how questions.

• Physical: A body the size of a 5-year-old boy.

Kenny has had stem-cell treatments in 2009, 2010, and May and November of 2011. The repeated treatments are required because adult stems cells will work repairing cells for a period of time, about six months, then leave the body.

“When I think I’ve seen his skills level out, we’ll go for another treatment,” explained Marty.

What are some of the changes that Kenny has undergone in the past three years? First came the ability to read and clearer speech.

“When he got back, he just picked up a book and started reading, and I could understand every word,” said Mike Hughes, Marty’s brother. “It was like a light just turned on.”

Other gains: Kenny is talking about past events for the first time, and he’s conversational now. He expresses opinions and looking ahead to the future. He was finally potty trained at age 9. He’s doing math now. He’s calmed down considerably. This summer, he went to summer camp, staying overnight for three nights, in the same cabin as Philip.

“There’s no doubt in my mind how much he’s progressing,” Marty said. “We’re working on catching up right now, and how do we best do that?”

The costly treatment, which isn’t covered by insurance, hasn’t been approved yet by the Food and Drug Administration. Despite the fact that the stem cells come from the human body, the cells are considered a new drug by the FDA and are subject to stringent research and testing that can take years.

So this leaves the Kelleys and others like them seeking stem-cell treatment, going overseas to get it.

“It’s just a matter of how much are you going to spend,” Marty said. “There’s no treatment here that was going to do this much for him.”

Thursday, September 13, 2012

Medistem Advances Type 1 Diabetes Stem Cell Technology Licensed From Yale

SAN DIEGO, CA -- (Marketwire) -- 09/12/12 -- Medistem Inc. (PINKSHEETS: MEDS) announced today completion of the first phase of a joint project with the Shumakov Research Center of Transplantology and Artificial Organs of the Russian Federation and its Russian and CIS licensee ERCell. The collaboration is based on using Endometrial Regenerative Cell (ERC) technology licensed from Yale University to treat type 1 diabetes.

Dr. Viktor Sevastianov, Head and Professor of the Institute of Biomedical Research and Technology, within the Shumakov Center, demonstrated safety and feasibility of ERC injection in experimental animal models of diabetes. Additionally, the studies demonstrated that the cell delivery technology developed by Dr. Sevastianov's laboratory can be used to enhance growth of ERC. These experiments are part of the process for registration of "new pharmacological substances," which is the first step towards drug approval in Russia.

"Type 1 diabetes is a significant problem in the Russian Federation. Our laboratory has been working developing various delivery formulations for cell therapy, such as SpheroGel, which is registered in Russia," said Dr. Sevastianov. "Given that the ERC can be produced in large quantities, is a universal donor cell, and already is approved for clinical trials in both the USA and Russia, we are optimistic our collaboration will lead to a viable commercial product for the type 1 diabetes Russian population."

Medistem discovered ERCs in 2007, and they appear to possess "universal donor" properties, allowing the cells derived from one donor can treat multiple unrelated recipients. According to Medistem's current FDA cleared production scheme, one donor can generate 20,000 patient doses. Medistem licensed technology from Yale University for generating insulin producing cells from ERC. A publication describing the technology may be found at http://www.ncbi.nlm.nih.gov/pubmed/21878900.

"Our vision is to combine SpheroGel, which is a clinically-available cell delivery vehicle in Russia, together with Medistem's ERC and technology from Yale University to generate a commercially-viable product for clinical trials in type 1 diabetes patients," said Thomas Ichim, CEO of Medistem.

Medistem has outlicensed the Russian and CIS rights to ERC and related products to ERCell LLC, a St. Petersburg-based biotechnology company. Under the agreement, Medistem owns all data generated and will receive milestone and royalty payments.
"By working with leading investigators in Russia and the USA, we seek to be the leaders in a new era of medicine in Russia," said Tereza Ustimova, CEO of ERCell."

Cautionary Statement This press release does not constitute an offer to sell or a solicitation of an offer to buy any of our securities. This press release may contain certain forward-looking statements within the meaning of Section 27A of the Securities Act of 1933, as amended, and Section 21E of the Securities Exchange Act of 1934, as amended. Forward-looking statements are inherently subject to risks and uncertainties, some of which cannot be predicted or quantified. Future events and actual results could differ materially from those set forth in, contemplated by, or underlying the forward-looking information. Factors which may cause actual results to differ from our forward-looking statements are discussed in our Form 10-K for the year ended December 31, 2007 as filed with the Securities and Exchange Commission.

Contact: Thomas Ichim Chief Executive Officer Medistem Inc. 9255 Towne Centre Drive Suite 450 San Diego, CA 92122 858 349 3617 www.medisteminc.com twitter: @thomasichim
Source: Medistem Inc.

Monday, September 10, 2012

Adipose (Fat) Stem Cell Counting Methods Can Lead to Inaccurate Dosing

David G. Morrison, Dirk A Hunt, Isaac Garza, Robbie A. Johnson, Mary Pat Moyer*
INCELL Corporation LLC www.incell.com
12734 Cimarron Path, San Antonio, TX 78249

*Corresponding Author Contact information: mpmoyer@incell.com; 210.877.0100

INTRODUCTION

There has been considerable interest in stem cell treatment of humans and animals for osteoarthritis and other conditions in recent years. Quality and accuracy of the methods of isolation and counting of cells for therapeutic dosing is of great concern to practitioners whether their patients have two or four legs. INCELL Corporation is a GMP cell therapy manufacturer of human tissue derived cells and has extensive experience isolating stem and stromal vascular cells from fat removed from humans and many animal species.

INCELL is actively evaluating process methods to improve quality and quantity of cell therapy preparations, such as mammalian stromal vascular fraction (SVF) including stem cells isolated from adipose tissues. Intrigued by the high cell numbers (5 to 20 million cells/gram1-3) reported by kit/device manufacturers such as MediVet-America (Lexington, KY), Intellicell Biosciences (New York, NY), and Adistem, Ltd. (Hong Kong) in adipose stem cell therapy compared to other methods (e.g., 4-6), INCELL staff conducted a research study to investigate the high apparent yield of stem cells. This initial work was focused on SVF cells from the MediVet Kit, which is marketed to isolate adipose-derived canine SVF and stem cells.

The cell yields reported for the Medivet Kits are five to more than ten times higher than the yields routinely obtained by INCELL from freshly harvested human or animal adipose tissue using our adipose tissue processing methods. These yields are also tenfold or higher than those reported in the literature by most academic researchers (Chung-canine4, Vidal–equine5, Yoshimura–human6). Since these cell counts are used to support stem cell dosing recommendations and cell banking, it is important to better understand why the cell numbers are higher. If the numbers are accurate, this would be process improvement, but if they are inaccurate, then there is a risk of incorrect cell numbers that will impact therapeutic dosing by the practitioner and/or biorepository storage. Particularly important are the numbers of renewable, replicating cells that grow out in culture as attached fibroblastoid (F) cells and are quantified colony-forming units (CFU-F). When recommended processing methods and commonly used counting methodologies are compared to CFU-F results from the same processed tissue counting errors are evidenced as differences in CFU-F numbers.

A comparative analytical study of three dog donors of adipose tissue was designed to evaluate the cell yields using the MediVet Kit as an example of this class of isolation system. All kit procedures were followed as per the instructions provided. A brief overview of the different cell counting methods used, and the resultant cell counts, observations and explanations of the results observed, are described below.
INCELL White Paper: Stem Cell Counting Methods Can Lead to Inaccurate Dosing


METHODS AND RESULTS

The Cellometer (Nexcelom Biosciences, Lawrence, MA) is commonly used for cell counting and is recommended for use by MediVet. It uses a two-part dye that counts by staining DNA in live cells green with acridine orange (AO) and dead cells red with propidium iodide (PI) at the same time. The problem with using AO staining as an indicator of living cells is that background lipid micelles auto-fluoresce green and would be detected as AO stained cells. Emulsifying agents used in these methods (such as Solution E in the kit), when mixed with water, can form a myriad of small fat droplets, called micelles or liposomes. Figure 1 shows that kit-recommended settings on the Cellometer overestimated mean SVF live cell counts compared to other methods investigated. The higher Cellometer counts are at least partially explained by the machine counting the micelles. As part of the reagent cross-check, Solution E (emulsifying agent) from the tested kits was evaluated without adding any adipose tissue or SVF. The Cellometer counted the micelles as “cells” (Figures 1 and 2), leading to erroneous cell counts due to the Solution E micellular structures. In order to completely differentiate these background lipid particles from live cells in any cell therapy product, methods to specifically identify cell nuclei are required.

Figure 1. Mean Nucleated Cell Counts per Gram of Adipose Tissue


Legend: Mean (+ SD) nucleated cell counts per gram of adipose tissue are shown for four different counting methods from 3 dogs, with counts in triplicate. Additionally, the Cellometer count for Solution E without any cells is shown. The Solution E count accounts for nearly 75% of the inflated Cellometer SVF counts and demonstrate that most counts shown by the Cellometer are not cells. When the NucleoCounter was used to count Solution E (data not shown), it found no cells because of a lack of any nuclei to stain with PI. Using the Cellometer with the AO dead/alive program recommended by MediVet, the machine counted micelle bodies as cells because it was unable to differentiate between the green autofluorescence of the micelles and the green AO staining that would have been present if there were any cells in the sample. Overall, the Cellometer reported a cell count approximately 5 to 16 times higher than the manual cell counts.

Coulter-type Counters. An alternative method recommended in the instructions to the kit is cell counting on a clinical hematology analyzer. Coulter counter methods measure the electrical impedance as cells pass between electrodes. Cells and micelles would have similar impedance, leading to the high cell count seen in clinical Coulter-type counters. A Heska HemaTrue counter was evaluated in this study (Figure 1). That work was done off-site and immediately at a local veterinary practice within 0.2 miles of the INCELL offices.

NucleoCounter. A common automated counter used with SVF cells is the NucleoCounter (ChemoMetec; Denmark). This counting machine and method is based on staining cell nuclei with PI dye. The NucleoCounter uses a two-stage process to achieve a viable SVF cell count, and counts are not greatly affected by the presence of micelles. The cell counts from the NucleoCounter are more in line with the manual hemocytometer counts and more closely reflect the outgrowth of live cells in culture as colony-forming units (CFU below). The NucleoCounter method has been reported in the literature by human adipose stem cell companies (e.g. Cytori, San Diego, CA) and by at least one veterinary stem cell company (Vet-Stem, Inc., Poway, CA) as being an accurate automated counting method for SVF cells.

Hemocytometer. SVF cells were also counted manually using a hemocytometer with a combination of DAPI (blue wavelength fluorescent nuclear stain) and trypan Blue (dye excluded by viable cells) staining by overlaying the digital light images and DAPI fluorescent images taken on a fluorescent microscope and counting the DAPI-stained nuclei in cells that excluded Trypan Blue (Figures 1 and 2). This provides an approximation of the “true” nucleated, live cells in a population. This is the method used to visually reduce errors of counting non-cellular materials as cells. However, it requires specialized expertise and training, and an imaging fluorescence microscope to clearly distinguish cells. It also takes considerable time for each sample.

Figure 2. Micrograph of Isolated SVF cells by the MediVet Method


Legend: This photomicrograph (bar=200 microns) shows the final SVF preparation from the MediVet kit process and what looks like a dense covering of cells. most of the small cell-like structures, however, are lipid droplets or micelles. These are carried into the cell preparation, have green autofluorescence and appear to the Cellometer as countable units, which may explain why the reported cell numbers from this method are higher than other industry or academic reports.


Colony-Forming Unit Assay (CFU). assays were also done as a measure of the number of stem cells in the population. In this assay, a defined number of SVF cells were placed in culture plates and allowed to attach and grow into visual colonies (also known as a “CFU-F” assay). While only a subset of viable cells in the SVF will attach to the plastic dishes and form colonies, the CFU assay provides a good index for proliferative potential. Thus, the outgrowth of renewable stem cells as measured by CFUs comprises a fraction of the population. As expected, there was variability between animals and counting methods. The highest percentage was 8 to 10% of the cells seeded in the NucleoCounter counts and in all cases the Cellometer CFUs were lower for each dog tissue donor and overall (Figure 3). The differences in CFUs between methods were statistically significantly (p<0.05) in all animals and in the overall composite results comparing the NucleoCounter and the Cellometer. The lower CFU observations did not correlate with higher numbers of cells (Figure 1). This led to the conclusion that the cell numbers in the Cellometer are an overestimate, since the source tissue processed SVFs which were used for the counts are the same.

Figure 3. Cell Counting Methods and CFU-F of Seeded Cells


Legend: SVF cells were obtained for dog donor fat samples (N=3; 3 replicates) processed according to Medivet kit instructions. The resultant SVF cells were counted by a variety of methods. Cells were seeded into complete MSC culture medium for CFU studies with cell numbers for seeding designated according to two of the counting methods used: NucleoCounter (blue) and Cellometer (brown). After the colonies formed, they were counted and calculations were done to determine the Mean +/- SEM numbers of CFU-F per 200,000 cells seeded. These data were calculated for each individual dog and for All pooled data with each counting method. Statistical analyses (Students t tests for NucleoCounter vs. Cellometer) between the 2 counting methods showed statistical significance at p<0.05 for the individual dogs and overall.

Light Activation. As part of evaluating process improvements, Platelet Rich Plasma (PRP) in concert with the Medivet light activation, as per the kit instructions, was used to assess the increase of cell viability and the ability to form colonies in CFU assays as reported by MediVet1. Figure 4 results clearly show that light activation did not increase the number of stem cells or increase their ability to proliferate as measured by CFU assays in this study. In fact, 2/3 dog donors showed significant reductions in percent CFU after the exposure to the light activation system, whereas Dog #2 was essentially unchanged (Figure 4). Reasons for the variability and this unexpected result were not investigated further.

Figure 4. Colony Forming Units and Light Activation


Legend: Percent CFU-F assays show estimates of stem cell content/activity from a sample taken before (left columns) and a sample taken after (right columns) the MediVet light activation step. Data show that exposure to the light activation system resulted in a reduction in the mean number of colonies in Dogs #1 and #3 and no real difference in Dog #2. The average reduction in %CFU across all groups was approximately 34%.

SUMMARY
This study shows that incorrect counting of adipose derived SVF cells and the subset of regenerative stem cells can subsequently result in inaccurate dosing, both in direct therapeutic applications and in cryostorage of cells for future use. The DAPI-hemocytometer cell count (manual) was considered the most accurate, but there are various sources of technical difficulties that can lead to incorrect cell numbers. The nature of adipose tissue itself with variability in dissociation by enzymatic digestion can all contribute to the outcomes. Fat tissue has a propensity to form acellular micelles and oils upon tissue disruption. Processing methods or reagents (e.g., Solution E or lecithins) can generate micelles that may be erroneously counted as cells. Autofluorescence and dye trapping or uptake by the micelles can lead to very high inaccurate cell counts when automated cell counting is used.

In this study the most inaccurate counting came from the Cellometer. When used according to kit-recommended guidelines and on-site training provided by Nexelcom for counting cells by the MediVet procedure, the Cellometer overstated the DAPI-hemocytometer cell count by up to 20X or more. The Coulter Counter protocols also led to incorrect, high cell numbers. Although the cell counts were still a bit high, the authors recommend the NucleoCounter, or similar equipment, as more acceptable for automated counting. The manual hemocytometer-DAPI method is the most accurate, but requires a highly experienced cell biologist or technician to make accurate counts and is not suitable for routine clinical use.

The SVF was shared among the counting and test methods. Thus, the post-processing numbers of stem cells would be the same and cell numbers should directly correlate with CFU-F per cell numbers seeded if the cell counts are accurate. Lower CFU numbers than expected from the cell counts would directly demonstrate an error in the cell counting method. To that end, the CFU-F results for the NucleoCounter showed 2.5X to about 20X higher CFU-F than the Cellometer leading to the conclusion that the Cellometer counts are high and incorrect. Also, in this study the percent CFU was reduced or there was no change after light activation, demonstrating no benefit or a detrimental effect of this step. Significantly fewer cells suggested that a death pathway may have been induced by the light treatment in 2 of 3 dog donors, but further studies are needed to clarify the proposed mechanisms of action and the controlling factors of the outcomes. These might include clinically relevant features of the donor dogs, or equipment or technical issues.

Other companies also have claims of very high cell numbers when their processes are used. Adistem2, like MediVet1, states they add an emulsifying agent to their kits to assist in cell release, and they also use a light activation system. Their kits were not tested in this study but it is possible that the high cell numbers reported by Adistem are also incorrect and result from the same problems highlighted in this paper for the MediVet procedure. Ultrasonic energy, which is commonly used to manufacture micellular liposome structures and to disrupt and lyse cells, is another potentially problematic procedure for counting and verifying viable, regenerative cells. Intellicell3 uses ultrasonic energy to release cells from adipose tissue, and it is possible that resultant micelles or cell fragments contribute to the higher than expected cell numbers. This assumption could be verified with additional studies.

In summary, the authors caution that great care must be taken when using kits and automated cell counting for stem cell dosing and cryobanking of cells intended for clinical use. Overestimated cell numbers would be a major confounding source of variation when efficacy of stem cells injected are compared as doses based on cell number and when cryostored cells are aliquoted for use based on specific cell numbers as a treatment dose. Hopefully, this study will lead to more reproducible counting and processing methods being reported in the literature, more inter-study comparability of cell doses to clinical outcomes, more industry diligence to support claims, and more accurate counting for dosing stem cell therapies to patients.

REFERENCES
1 http://www.MediVetlabs.com/cellcounts.html; accessed June 21, 2012.
2 http://www.adistem.com/science-and-technology.htm
3 http://www.intellicellbiosciences.com/intellicell-facts.html
4 Chung D, Hayashi K, Toupadakis A, et al. Osteogenic proliferation and differentiation of canine bone marrow and adipose tissue derived mesenchymal stromal cells and the influence of hypoxia. Res Vet Sci, 2010; 92(1):66-75.
5 Vidal MA, Kilroy GE, Lopez MJ, Johnson JR, Moore RM, Gimble JM. Characterization of equine adipose tissue-derived stromal cells: adipogenic and osteogenic capacity and comparison with bone marrow-derived mesenchymal stromal cells. Vet Surg, 2007; 36:613–622
6 Yoshimura K, Shigeura T, Matsumoto D, et al: Characterization of freshly isolated and cultured cells derived from the fatty and fluid portions of liposuction aspirate. J Cell Phys, 2006; 205:64-76.

Disclosures
This study was performed as independent research by INCELL staff, but was funded in part by Personalized Stem Cells, Inc., Ramona, CA and VetStem, Inc. INCELL is not involved in the clinical veterinary stem cell business, but does provide services for veterinary and human R&D, contract manufacturing, and stem cell services to the human stem cell industry.

Medistem Panama - Inside our adult stem cell laboratory || Video




• Learn how cord blood stem cells, cord tissue stem cells and stromal vascular fraction cells (fat stem cells and T-regulatory cells) are processed in our GMP and GLP compliant lab.

• See inside our 3 class 10,000 clean rooms and 8 class 100 laminar flow hoods.

• Discover what mesenchymal stem cells, CD34+ stem cells and endometrial stem cells look like under the microscope.

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