• Defining a stem cell product – working proposal and recommendations
    Last 2 months I was discussing a possible definition of “stem cell product“. I polled professionals and had few opinions exchange via email and LinkedIn. Today, I’m going to summarize this discussion and propose a working definition and recommendations. Why do we need to define it? 93% of polled professionals think that it is important issue to discuss. 7% think that there is no point to do it. Unfortunately, both sides did not elaborate publicly – why they think so. […]
  • Methods and Protocols Digest – July, 2014

    by Alexey Bersenev on July 24, 2014 · 0 comments

    in methods,protocols

    Methods and Protocols Digest is a collection of links to the methodological articles, freely available online. Please share your protocol with community!

    FEATURED:

    Reprogramming
    Direct reprogramming skin fibroblasts of diabetic patients in insulin+ clusters (PLoS ONE)
    Direct reprogramming of mouse fibroblasts to neuronal cells (Sci World J)

    Mesenchymal Stromal Cells
    Characterization of 4 different populations of human MSCs (Int J Mol Med)
    Clonal analysis of human CD271+ MSCs (Hematologica)
    Method for 3D expansion of human bone marrow MSC (PLoS ONE)

    Pluripotent stem cells
    Human ES cell-derived insulin+ cells in diabetes model (PLoS ONE)
    Depletion of residual pluripotent cells in iPS differentiation culture by inhibition of DNA topoisomerase II (Stem Cells Dev)
    Generation of cardiomyocytes from pluripotent stem cells in suspended microcarrier cultures (Stem Cell Res)
    Robust enrichment of cardiomyocytes derived from human pluripotent stem cells (Stem Cell Reports)
    Generation of oligodendrocytes from multiple sclerosis patients by iPS (Stem Cell Reports)

    Endothelial cells
    Generation of endothelial cells from peripheral blood (J Cell Mol Med)

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    20 bioprinting companies to watch

    by Alexey Bersenev on July 23, 2014 · 0 comments

    in notes

    Bioprinting is a new buzz word in biomedicine. With creation of new 3D printers and increased interest from investors, the field is starting to gain momentum and commercialize. But if you look at bioprinting as a business, all you will see is news about Organovo. But, in the last few years many new bioprinting companies are popped up. In this post I’ll highlight 20 most notable companies, developing bioprinting products.

    1. Cyfuse Biomedical
    Japanese company, founded in 2010. Makes bioprinter Regenova. Working on bioprinting few organs and tissues for drug discovery and regenerative medicine.

    2. TeVido Biodevices
    US-based private start-up company, founded in 2011. The company is working on printing breast tissue with nipples and cellular wound cover, created from patient’s adipose stem cells. Learn more from this video.

    3. Regenovo Biotechnology
    Chinese company, founded in 2013. Commercializing bioprinter “Regenovo” for “making biomaterials, organs and tissues” for regenerative medicine.

    4. Aspect Biosystems
    Private Canadian company, founded in 2013. Commercializing bioprinted “living human tissues on demand” – for Pharma R&D.

    5. Rainbow Biosciences
    A division of US public company Rainbow Coral Corp. Since 2012, the company has a joint venture and commercializing product, developed by nano 3D Biosciences – 3D BiO Assay. This assays is claimed as “the first commercially available 3D bioprinting system for drug screen”. The technology behind – magnetic levitation.

    6. RegenHU
    Private Swiss company, founded in 2007. Has few bioprinting products on a market – printer BioFactory and matrix BioInk.

    7. Osteopore International
    Clinical stage company from Singapore. Printing matrices for bone regeneration. Has 2 products on a market – Osteoplug and Osteomesh. For matrix fabrication company uses QuickSlice software offered by Stratasys Inc.

    8. Bio 3D Technologies
    Private Singaporean company, founded in 2013. Makes bioprinter – Life-Printer “X” for Pharma R&D and potential RegenMed applications.

    9. GeSiM
    German company, which makes equipment for liquid handling automation and microfluidics platforms. The company makes and sells bioprinter BioScaffolder 2.1 for printing of scaffolds in tissue engineering.

    10. Tissue Regeneration Systems
    Private US company, founded in 2008. The company “fabricate skeletal reconstruction implants”. Last year, FDA cleared the first bioprinted implant for bone regeneration as 501k device.

    11. nScrypt Inc
    US company, founded in 2002. Recently nScrypt is commercializing products for tissue bioprinting – Computer Aided Machining equipment and biomaterials.

    12. OxSyBio
    U of Oxford spin-out, founded in 2014. Company will develop bioprinted materials for wound healing and drug delivery. Read more.

    13. Next 21
    Japanese company, which provides a service for printing of custom bone implants. The company filed application for approval of bioprinter.

    15. DigiLab Inc
    US company, which makes variety of devices for life sciences. The recent product CellJet – automated live cell printer for research applications.

    16. Advanced Solutions Life Sciences
    A division of Advanced Solutions, dedicated to development software and hardware for 3D printing of tissues. Commercializing two products – Tissue Structure Information Modeling (TSIM) and BioAssemblyBot.

    17. MicroFab Technologies Inc
    US company, which offers commercial printing platform JetLab. The platform could include custom TissueJet bioprinter for tissue engineering.

    18. EnvisionTEC
    International 3D printing industry giant. A big part of company now dedicated to biofabrication. Company makes and sells printer 3D-Bioplotter for applications in tissue engineering.

    19. Seraph Robotics
    Private US company, founded in 2011. Company makes and sells 3D printing platform (hardware and sofware) Fab@Home Model 3. This platform successfully used in academic institutions to print scaffolds for tissue engineering.

    20. Organovo
    Public American company – the icon of bioprinting business. Makes printer Novogen MMX and provide tissues (NovoTissues) for R&D.

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    Cells Weekly – July 20, 2014

    by Alexey Bersenev on July 20, 2014 · 0 comments

    in notes

    Cells Weekly is a digest of the most interesting news and events in stem cell research, cell therapy and regenerative medicine. Cells Weekly is posted every Sunday night!

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    1. Criticism of “cord blood for autism” trial
    Criticism of recently launched autism trial at Duke U is keep coming. SFARI posted a big piece about this trial, citing stem cell scientist Arnold Kriegstein:

    “It’s probably premature to run large trials without evidence that they have a therapeutic effect that [we] understand,” cautions Arnold Kriegstein…
    “These are not cells that can treat a laundry list of diseases,” he says. Because the stem cells are similar to those that normally give rise to blood cells, he says, it is unlikely that they can repair or replace neurons in the brain. Also, because autism results from errors during development, it is unlikely that the stem cells can reverse those effects…

    It was re-posted by Scientific American and went mainstream. Many stem cell researchers agree with Kriegstein, for example Jeanne Loring:

    I side with Arnold Kriegstein on this issue, and am glad that he is willing to be so straightforward.

    I’m afraid that studies like these are used as rationalization for charlatans.

    I don’t see the reason for such “hysteria”. Many cell therapy trials, approved by regulatory agencies, lack of strong scientific background. This one is not an exclusion, but, at least has a rationale. Many human conditions have no good animal models and the way to test it is via approval for first-in-human trial. And what can we learn for such criticism? Do you remember the same kind of criticism for same kind of US-based trial 2 years ago? It’s over and over again. Was this criticism valuable? What did we learn from it?

    2. Whistleblower is suing stem cell company
    This week, we witnessed unique case in cell therapy history – former employee sues stem cell therapy company:

    Rob Williams sued StemCells Inc. on Monday in Alameda County Court, alleging wrongful termination, retaliation and violation of the California False Claims Act.

    Apparently, he saw a lot of deficiencies in stem cell product manufacturing, which are not in compliance with GMP rules and put patients safety at risk:

    “Shortly after beginning his employment, plaintiff noted poor sterile technique, failure to adhere to current Good Manufacturing Practices in the company’s manufacturing process, and substantial deficiencies in the company’s Manual Aseptic Processing of HuCNS-SC (Human Central Nervous System Stem Cells) cell lines – failure and deficiencies that put patients at risk of infection or death during ongoing clinical trials,” Williams says.

    After failed attempts to fix these deficiencies by reporting to upper management, he was fired by company. California Stem Cell Report blog nicely followed this story here, here and here. Company’s representative responded via email to California Stem Cell Report:

    “The Company has reviewed the complaint filed by Mr. Williams, a former employee whose employment was terminated for performance deficiencies, and finds no merit to the allegations…”

    As far as I know, this is the first scandalous whistleblower-related case in the history of cell therapy.

    3. Current state and controversies in cardiac cell therapy
    In relation to recent controversy around cardiac stem cells and Anversa’s lab, good critical analysis have been recently written. First, I’d like to highlight excellent blog post from Jalees Rehman – The Road to Bad Science Is Paved with Obedience and Secrecy. He went through history of discoveries and controversies, surrounding cardiac cell therapy:

    Considering the size of the Anversa group – consisting of 20, 30 or even more PhD students, postdoctoral fellows and senior scientists – it is puzzling why the discussions among the group members did not already internally challenge their hypotheses and findings, especially in light of the fact that they knew extramural scientists were having difficulties replicating the work.

    Another good piece, was published in the last issue of Science and entitled “The elusive heart fix” (unfortunately, behind paywall). This piece is full of “lessons to learn” and great quotes from professionals in the field. Some of my favorite quotes:

    Charles Murry: “Everybody has wanted to be the first toregeneratetheheart.”

    Cardiologists acknowledge that cell therapy was marked by outsize expec- tations. “We all hype our work,” says Steven Houser, a cardiac muscle biologist at Temple University… “the hype got ahead of the science.”

    …scientists publish positive results far more often than negative ones, and that may imply that a novel treatment is more promising than it really is.

    … the science is in a state of flux. The idea that bone marrow cells morph into heart muscle has been largely abandoned.

    The trial, called BAMI, is “unethical,” Arnesen charges, because, he believes, the therapy it’s testing has already been shown not to help.

    I’d highly recommend you to read both – Rehman’s post and Science piece!

    4. Somatic in vivo reprogramming makes pacemaker cells
    The most interesting scientific paper of the week came up in Science Translational Medicine. Researchers used one transcription factor to reprogram cardiomyocytes in peacemaker cells in pig model of complete heart blockade:

    Thus, minimally invasive TBX18 gene transfer creates physiologically relevant pacemaker activity in complete heart block, providing evidence for therapeutic somatic reprogramming in a clinically relevant disease model.

    The Scientist nicely covered this paper:

    The team monitored the pigs for two weeks after injections and found that the activity of the induced pacemaker cells peaked at day eight and then slowly declined. This was not a surprise because adenovirus-infected cells tend to be cleared from the body, Cingolani explained.

    Such short-term reprogramming would be fine for patients requiring a temporary alternative to electronic devices, such as those undergoing treatment for pacemaker-related infections. Indeed, Cingolani said that while developing the technique his team’s aim was “to focus on these high-risk patients.” But for long-term reprogramming, an alternative vector would be necessary.

    5. Talk show on experimental stem cell treatment procedures
    Australian media outlet SBS recorded talk show about experimental stem cell treatments. I was really enjoying watching it, because there were all kind of people and opinions. Highly recommend you to watch!

    6. HIV stem cell chronicle – Berlin Patient – Boston Patients – Australian Patients
    Two new cases of “HIV clearance” by bone marrow transplantation were reported this week on 20th International AIDS Conference:

    The two patients, both Australian men, became apparently HIV-free after receiving stem cells to treat cancer. They are still on antiretroviral therapy (ART) “as a precaution”, but those drugs alone could not be responsible for bringing the virus to such low levels, says David Cooper, director of the Kirby Institute at the University of New South Wales in Sydney, who led the discovery.

    Even though mass media used term “cured”, we all know that it could be too premature. Remember HIV return in “Boston Patients” and “Missiscipi Baby”:

    Unfortunately, several months after the ‘Boston’ patients stopped taking ART, the virus returned. An infant born with HIV in Mississippi who received antiretroviral therapy soon after birth, then stopped it for more than three years, was thought to have been cured, but has had the virus rebound, too.

    7. The story of creation bioengineered blood vessels
    Scientific resource Nautilus posted very nice story about creation of bioartificial organs, taking blood vessels as example:

    This year, Niklason’s hollow, milky-white blood vessels are being implanted in patients enrolled in clinical trials in Poland and the United States. The first stage of testing will determine the vessels’ safety. The next stage of trials will demonstrate whether the tubes represent an improvement over existing techniques.

    Highly recommended!

    8. Fresh reviews:
    The Billion Cell Construct: Will Three-Dimensional Printing Get Us There?
    Injectable Biomaterials for the Treatment of Myocardial Infarction and Peripheral Artery Disease
    Sca-1+ Cardiac Progenitor Cells and Heart-Making: A Critical Synopsis

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    bonus:

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    I’ve written about aneuploidy as phenomenon, associated with expansion of mesenchymal stromal cells (MSC) and some other types of cells. Even though, aneuploidy is chromosomal abnormality, it is not necessarily associated with malignant transformation. It could “represent” senescence. The big question is how can we ensure safety of MSC-based clinical products with detected aneuploidy or other chromosomal abnormalities?

    Recently, a group of researchers from S. Korea, proposes to set a cut-off value for aneuploidy in clinical MSC cultures. They analyzed 103 cultures of 68 MSC lines, derived from different tissue sources:

    Interphase FISH showed variable aneuploid clone proportions (1% to 20%) in 68 MSCs. The aneuploidy patterns were asymmetric, and aneuploidy of chromosomes 16, 17, 18, and X occurred most frequently. Clones with polysomy were significantly more abundant than those with monosomy. The cut-off value of maximum polysomy rates (upper 95th percentile value) was 13.0%. By G-banding, 5 of the 61 MSCs presented clonal chromosomal aberrations. Aneuploidy was asymmetric in the malignant hematological diseases, while it was symmetric in the benign hematological diseases. We suggest an aneuploidy cut-off value of 13%, and FISH for aneuploidy of chromosomes 16, 17, 18, and X would be informative to evaluate the genetic stability of MSCs.

    Yet another recent study indicates to MSC clinical expansion-related chromosomal instability as very frequent phenomenon:

    Signs of chromosomal instability were observed in 11 out of 21 patients (52%). From a total of 910 analyzed metaphases, five chromatid gaps, six chromatid breaks and 14 tetraploid cells were detected giving as total of 25 metaphases with chromosome damage (2.75%).

    >50% of MSCs cultures on 2nd passage!

    How do you think these data should be interpreted? Should we be scary of these types of MSC chromosomal abnormalities? Is G-banding a method of choice? Is anueploidy ok for clinical culture and how much is ok? Would you agree with proposed threshold?

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    Impact of controlled-rate freezing interruption on cell viability

    by Alexey Bersenev July 16, 2014 clinical lab

    If you process cells for clinical use, you probably perform cryopreservation in the device, called controlled-rate freezer (CRF). This is the most frequent and routine procedure in cord blood banking. CRF uses liquid nitrogen (LN) to cool cells in controlled manner. The whole procedure takes usually 40-60 min and finished by freezing cells at -80C. However, sometimes such freezing procedure could be interrupted by errors (the most frequent cause – not enough LN or pressure in the tank, connected to […]

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    Cells Weekly – July 13, 2014

    by Alexey Bersenev July 14, 2014 notes

    Cells Weekly is a digest of the most interesting news and events in stem cell research, cell therapy and regenerative medicine. Cells Weekly is posted every Sunday night! 1. “Stem cell therapy” complicated by excessive tissue growth 8 years later Clinical case, described by Clare Wilson in New Scientist made a biggest buzz this week. A woman received autologous olfactory mucosal cell implantation for experimental treatment of spinal cord injury. Intervention was useless and 8 years later she developed back […]

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    Breaking down fat: Considerations for adipose tissue processing

    by Alexey Bersenev July 12, 2014 adipose

    Quality and therapeutic potency of stromal vascular fraction (SVF) is very much depending on processing of adipose tissue. Cell composition of SVF can vary dramatically between different methods of processing. Unlike SVF, adipose-derived stem cells (ASC) in culture could be more homogenous. Different adipose tissue isolation techniques could affect a gene expression profile of ASC. In this post, I’ll summarize some considerations in choosing the method of fat tissue processing for further SVF or ASC isolation. Fat tissue harvesting procedure. […]

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    Six reasons to work in academic cell manufacturing facility

    by Alexey Bersenev July 8, 2014 RegenMed digest

    Cell therapy field is booming and continue to offer new jobs. Many of you looking for new positions and facing a choice between industry and academia. Today, I’d like to highlight some key differences and advantages of working in academic cell manufacturing facility as compared to industry. These career tips based on my own experience. I hope, it will help to navigate you in career choice. Possibility to participate in many different clinical trials. Usually, cell therapy academic facilities support […]

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    Cells Weekly – July 6, 2014

    by Alexey Bersenev July 6, 2014 notes

    Cells Weekly is a digest of the most interesting news and events in stem cell research, cell therapy and regenerative medicine. Cells Weekly is posted every Sunday night! 1. STAP news This week, both STAP papers were finally officially retracted by Nature. It took 5 months! Here is my pick of the news: Scientists comment against RIKEN CDB dismantling Don’t Penalize All of RIKEN CDB for STAP Cell Flap Nature to pull STAP papers Obokata returned to RIKEN to conduct […]

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    Concentration of DMSO in clinical cryopreserved cell products

    by Alexey Bersenev July 2, 2014 cell product

    DMSO is the most frequently used cell cryoprotectant in research and clinic. However, there is no consensus on its optimal concentration in cellular products. Defining the right concentration and threshold is important, because DMSO toxicity can cause severe adverse reactions. The recent trend is reducing DMSO concentration and/ or replacement DMSO by other non-toxic cryoprotectors. Historically, 10% of DMSO is widely used for cell cryopreservartion. Many “cell therapy” centers follow this “standard”, but some have started to test lower DMSO […]

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