• Cryopreservation of mesenchymal stromal cells can attenuate clinical immune effects
    As Jacques Galipeau reported in conferences and in the paper, cryopreservation could negatively affect therapeutic “immunomodulatory value” of mesenchymal stromal cells (MSC). There was no independent confirmation of Galipeau’s findings, and many MSC product developers remained skeptical. This week, Katarina Le Blanc published a report, which supports Galipeau’s conclusions and provides more insight into potential clinical value of this phenomenon. Let me just say – this paper could change the field! Le Blanc concluded that freeze-thawed human MSC compared to […]
  • Cells Weekly – April 26, 2015

    by Alexey Bersenev on April 26, 2015 · 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. Opinions on human germline genome editing
    The discussion of the week is genome modification of human germline. A new wave of debate was triggered by recent study, which demonstrated possibility of genome editing on human embryos. Actually Nature News pulled the trigger, but not paper itself. Here I collected some interesting coverage from this week with variety of opinions:

    Genomes of human embryos edited for the first time
    (Genetic Expert News Service)
    A Moratorium on Human Gene Editing to Treat Disease Is Critical (Rudolf Jaenisch for Time magazine)
    Chinese paper on embryo engineering splits scientific community (ScienceInsider)
    ISSCR response to the publication of gene editing research in human embryos (ISSCR)
    CRISPR and the Amoral Othering of Chinese Researchers (Life as an Extreme Sport)
    Chinese Team Reports Gene-Editing Human Embryos (MIT Tech Review)

    2. Cardiac gene therapy trial failed in Phase II
    A year ago, US-based company Celladon got a “Breakthrough” designation by FDA for the first gene therapy drug candidate Mydicar. FDA made this decision, based on results of Phase 1 trials of Mydicar in heart failure. However, today, company reported a negative results of their Phase 2 trial CUPID2. More from Fierce Biotech:

    Celladon’s lead program crashed Sunday afternoon as the biotech ($CLDN) announced that its heart therapy Mydicar failed the primary and secondary endpoints in a Phase IIb trial, losing out in a full lineup of efficacy tests.
    At the time Celladon achieved the breakthrough designation last year, Zsebo told FierceBiotech that a Phase III followup could well be an expected requirement at the FDA, while the EMA might well go ahead with an approval based on positive Phase IIb CUPID-2 results. But that’s all academic now. At this juncture, the biotech has to figure out how to reorganize and push forward in the face of a stinging setback.

    Yet another gene therapy company Sangamo Biosciences reported termination of their Alzheimer’s disease program.

    Very disappointing week for gene therapy field! Will investors react?

    3. Role of endogenous retroviruses in human embryogenesis
    The most interesting discovery of the last year – endogenous retroviruses linked to pluripotency – got continuation in the new study from Stanford’s Joanna Wysocka:

    … HERVK is transcribed during normal human embryogenesis, beginning with embryonic genome activation at the eight-cell stage, continuing through the emergence of epiblast cells in preimplantation blastocysts, and ceasing during human embryonic stem cell derivation from blastocyst outgrowths. Remarkably, we detected HERVK viral-like particles and Gag proteins in human blastocysts, indicating that early human development proceeds in the presence of retroviral products.

    Read more from Stanford’s press release.

    4. Germline gene editing to eliminate mitochondrial disease
    Very interesting methodological approach for elimination of mitochondrial mutations was described this week in Cell. Researches used TALEN-based genome editing of germline (mouse) to get rid of mutated mitochondrial DNA. Even though, this technique can be an alternative to controversial 3-donor mitochondria transfer, it is still germline modification. Nature reported:

    Ocampo and Izpisua Belmonte say that they are in the process of acquiring discarded human eggs and embryos from a fertility clinic, and waiting for approval from an ethics board. They plan to develop a line of stem cells from these modified cells, but say that they will not implant embryos into mothers or allow them to grow.

    Also read coverage by Paul Knoepfler.

    5. Clinical updates
    One of the biggest ever reported cell therapy clinical study was recently published in Cell Transplantation. The authors used adipose tissue-derived stromal vascular fraction for treatment of osteoarthritis in 1114 (!) patients. They reported great efficacy of the treatment:

    At least 75% Score improvement was noticed in 63% of patients and at least 50% Score improvement was documented in 91% of patients 12 months after SVF cell therapy.

    Yet another study, published in the same journal, reports 3-year outcome after olfactory cell transplantation in spinal cord injury. The authors noticed some benefit, but not significant:

    The limited recovery suggests that OLP transplants alone do not have significant benefits but may provide a rationale for larger randomized trials or combination therapies.

    Finally, clinical case of MRI tracking autologous bone marrow MSC after intrathecal transplantation in spinal cord injury patient. There were no effects, but there were some complications:

    A focal hyposignal intensity of tagged bone marrow-derived stem cells was detected at his cervical spinal cord with magnetic resonance imaging at 48 hours, which faded after two weeks, and then disappeared after one month. No clinical improvement of the neurological function had occurred at the end of this study. However, at 48 hours after the transplantation, he presented with a fever, headache, myalgia and worsening of his motor function (by one grade of all key muscles by the American Spinal Injury Association impairment scale), which lasted for 48 hours.

    6. Scrutiny of transdifferentiation study on PubPeer
    This week, very interesting discussion happen on post-publication peer review platform PubPeer about paper: Generation of neural progenitor cells by chemical cocktails and hypoxia. Some issues with figures and conflicts of interests. Very interesting! Highly recommended!

    7. Using “stem cell label” for marketing
    Chris Centeno on his corporate blog brought up very interesting examples of using “stem cell label” for marketing by doctors. Very good read – highly recommended!

    A gentleman who was using the same fat graft that he had been using for years, but who had met with a regulatory attorney about the legality of calling it’s use a “stem cell face lift”. The attorney rightfully asked that if it wasn’t a stem cell procedure before, how could it be one now? The physician responded that he could sign up more patients that way.

    8. Cost-effectiveness of double unit cord blood transplantation
    Very good analysis by EuroCord on cost of double cord transplant was recently reported in Hematollogica:

    Our results showed that for adults with acute leukemia in first complete remission in France, double cord transplantation is more cost-effective than single cord blood transplantation, with better outcomes, including quality-adjusted life-years.

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    Genetic editing of human embryos

    by Alexey Bersenev on April 22, 2015 · 0 comments

    in gene transfer

    As you may have heard today from Nature News, Chinese researchers recently reported the first data on using CRISPR/Cas9 genome editing technique in human embryos. You can read study here (open access). The authors used “defective” tripronuclear zygotes, left over from IVF procedures. The study is short, but with a clear message – CRISPR/Cas9 human germline editing did not work the way it supposed to:

    • low efficiency of editing (~ 52%);
    • off-target genotoxicity;
    • unwanted mutagenesis;
    • genetic mosaicism.

    To me, the study looks good and valuable. I did not find anything unethical in it. Nature News reported that the study was submitted to Science/ Nature and was rejected, partly because of “ethical reasons”. Too bad for Science and Nature! I wonder who are these “ethical guards of Science and Nature”, who set standards for submitted manuscripts? This study is historic and opens a new chapter in CRISPR-based genome editing research. The only puzzling thing about it at the moment is ultra-rapid peer-review = 1 day. It looks very suspicious.
    Many commenters pointed out that this study is a signal to take actions toward moratorium on any human germline experiments with CRISPR (Yes, Sangamo’s guy and more). My response to it was voiced by wise Jen Doudna:

    There’s no way to unlearn what is learned. We can’t put this technology to bed. If a person has basic knowledge of molecular biology they can do it. It’s not realistic to think we can block it. Same thing with regulations. To imagine that we could have international regulations, it’s just not realistic, and in any case, how do you enforce them? I wouldn’t feel comfortable hiding away in the lab. The better path is to try to be open and transparent and to educate people who want to understand it. It’s such a wonderful technology in many ways.

    Also read good coverage by:
    Carl Zimmer
    Paul Knoepfler

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    Cells Weekly – April 19, 2015

    by Alexey Bersenev on April 20, 2015 · 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. Stem cells failed in stroke trial
    The biggest news of the week was a press release of US-based stem cell therapeutic company Athersys. Both – primary and secondary efficacy end points of their stroke trial were missed. Company’s CEO said:

    “While the trial did not achieve the primary or component secondary endpoints, we believe the evidence indicating that patients who received MultiStem treatment early appeared to exhibit meaningfully better recovery is very important and promising,” continued Dr. Van Bokkelen. “The results appear to confirm that our window of intervention with MultiStem therapy may extend well beyond the limits of current care.

    Despite some optimism of CEO, investors dump the company’s shares (dropped 35-50% on the day of announcement). Seem like the whole “cell therapy industry” sector was negatively affected by this news on Friday. It is definitely a big blow for the field. This is the second failed Athersys Phase 2 trial, supported by Pharma companies. Last April, Atherys with Pfizer reported failed results of ulcerartive colitis trial.

    2. Organ buds – a new way of generation mini-organs in vitro
    A group of Japanese researchers describes a new method for generation of self-assembling organoids – organ buds. The recipe for mini-organ is a mix of pluripotent stem cells, endothelial cells and MSC. The authors were able to generate multiple organ buds and successfully tested them in vivo:

    Here, we demonstrate a generalized method for organ bud formation from diverse tissues by combining pluripotent stem cell-derived tissue-specific progenitors or relevant tissue samples with endothelial cells and mesenchymal stem cells (MSCs). The MSCs initiated condensation within these heterotypic cell mixtures, which was dependent upon substrate matrix stiffness. Defining optimal mechanical properties promoted formation of 3D, transplantable organ buds from tissues including kidney, pancreas, intestine, heart, lung, and brain.

    This is the one of the most impressive methodological studies this year so far!

    3. More on CAR T-cell therapy in solid cancers
    CAR T-cell therapy is powerful tool for experimental treatment of some types of leukemia. However, the whole field is waiting any signals of potential efficacy from targeting solid cancers by CART. Results of the first CART-cell trial, released about a month ago were not impressive. Today, some news came up about Novartis/ Upenn data, presented at annual AACR meeting:

    No major adverse events associated with the infusion of CART-meso were reported in the five patients, although other serious side effects, including sepsis, shortness of breath and elevated white-blood-cell counts were reported during follow-up…
    CART-meso appears well-tolerated so far, but does it work? Anti-tumor efficacy in the study was “suggested” by the clearing of malignant cells in the fluid surrounding the lungs of a single patient, researchers said. There was also imaging and clinical evidence of a tumor stabilizing or shrinking in another patient.

    4. Gene therapy of SCID is better than stem cell transplant
    Hematopoietic stem cell transplant and cell-gene therapy are 2 potentially curative therapeutic modalities for kids with genetic immunodeficiency (SCID). For children with completely matched donor, stem cell transplant is a method of choice. However, if there is no matched donor, other alternatives should be pursued. The first unique study, recently published in Blood, compares outcomes of partially matched stem cell transplant versus gene therapy:

    Our results show a clear advantage in terms of T cell development of gene therapy over HSCT with a mismatched donor. Patients treated with gene therapy display a faster T cell reconstitution and a better long-term thymic output. Interestingly, this advantage of gene therapy versus haploidentical HSCT seems to be independent of the existence of clinical graft versus host disease in the latter condition.

    Also read the Stem Cellar post about this study.

    5. CRISPR patents fight is getting hotter
    MIT Tech Review covered recent news about new development in legal patent battle about CRISPR genome editing rights.

    In a request filed Monday, the regents of California’s public university system asked the patent agency to reconsider ten patents issued starting last year to the MIT/Harvard Broad Institute, in Cambridge, Massachusetts, saying the hugely valuable rights should belong to them.
    The CRISPR-Cas9 editing technology was publicly described in the journal Science in 2012 by Jennifer Doudna, a biologist at the University of California, Berkeley, and the French microbiologist Emmanuelle Charpentier. But Feng Zhang, a scientist at the Broad Institute, was first to win a patent on the technique after submitting lab notebooks he says prove he invented it first…

    Is this fight about “billion-dollar” opportunities or scientific authority?

    also read:
    Xconomy piece on CRISPR patent fight
    Interview with Jennifer Doudna by Paul Knoepfler

    educational video:

    6. Cell lines authentication – how to deal with a problem of cross-contamination
    Nature magazine posted new requirements for authors about authentication of cell lines. The problem of cell lines cross-contamination is not new, but did not get enough attention. This week, Nature published a paper, which describes measures and resources for cell line authentication and prevention of cross-contamination.

    7. Tissue engineering pioneer Paolo Macchiarini cleared in misconduct case
    A famous surgeon and tissue engineer Paolo Macchiarini was accused in research misconduct last year. Karolinska Institute, where Macchiarini holds a Professor title, started investigation of 2 cases.
    This week, ScienceInsider reports an update on Paolo Macchiarini’s cases investigation:

    The decision, announced today, was made on 7 April by the Karolinska Institute’s vice-chancellor, Anders Hamsten, on the basis of an internal investigation by the institute’s ethics council. The council concluded that the issues raised are of a “philosophy-of-science kind rather than of a research-ethical kind.”

    Pierre Delaere, a head and neck surgeon at UZ Leuven in Belgium who brought the case against Macchiarini, says he is “stunned about such outright injustice.”

    Well, one more case to go.

    8. More on Gordie Howe’s case
    There were a lot of discussions on controversial case of experimental stem cell treatment of sport celebrity Gordie Howe. The case is continue to unfold. Leigh Turner and Paul Knoepfler wrote good posts on Howe’s case here and here. I’d highly encourage you to read both. Stem cell company + celebrity + hype = red flag!

    9. New methods and protocols:
    Generation and characterization of BMP4-primed pluripotent stem cells (PNAS)
    Cardiac tissue engineering on recombinant spidroin fiber meshes (PLoS ONE)
    Characterization of long-term cryopreserved human adipose-derived stem cells (Sci Reports)
    Human epithelial enteroids and colonoids from whole tissue and biopsy (JoVE)
    Protocol for neural induction of bone marrow MSCs (Cell J)

    10. New reviews:
    Cellularization strategies for tissue engineering of whole organs (Frontiers in Bioeng Biotech)
    Challenges in identifying the best source of stem cells for cardiac regeneration therapy (Stem Cell Res Ther)
    Wharton’s Jelly Derived Mesenchymal Stem Cells: Future of Regenerative Medicine? (Biomed Res Int)

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    Products in the pipeline – StemCells huCNS-SCs

    by Alexey Bersenev on April 15, 2015 · 1 comment

    in cell product

    This is 5th post of the series Products in the Pipeline. In this series we (community!) will provide publicly available information about therapeutic products – candidates for commercialization by cell therapy industry companies.
    ________________________________________
    Prepared by m.cea for StemCellAssays.com, narrowed by Alexey Bersenev .

    Product candidate name: huCNS-SCs (no trade name yet)
    Developer: StemCells Inc.
    History of development: Since 1990 (NeuroSpheres Holdings Ltd, Canada)
    Type of cells: Allogeneic, unmatched human central nervous system (brain) stem cells (huCNS-SCs), expanded ex vivo.
    Tissue source: Fetal brain donated 2nd trimester (16-20 weeks) tissue, obtained from tissue procurement agency Advanced Bioscience Resources.
    Processing steps: Master cell bank (MCB): Dissection of donated human fetal brain tissue, enzymatic digestion, labeling with CD133 and CD24 antibodies, FACS sorting of CD133+CD24-/lo (BD Vantage), neurosphere suspension culture in a chemically defined,serum-free medium X-VIVO15 supplemented with N2, heparin, N-acetyl cysteine (NAC), FGF2, EGF and LIF) at a density of 105 cells/ml, harvest, cryopreservation at an early passages. Working cell banks are generated from the MCB and cryopreserved at later passages.
    Expansion: huCNS-SC passaged rate 6>10+
    Phenotypic composition: CD133+, CD34-, CD45-, CD24-/lo, Nestin+, Sox2+
    Stability/Safety: Genetically & morphologically stable, normal karyotype, normal senescence
    Proposed mechanisms of action in respective indications:

    •  Neuronal Ceroid Lipofuscinosis (NCL or “Batten Disease”): Cell transplant for cross-correction of enzyme deficiency through donor cell secreted PPT-1 and TPP1 for host neurons absorption via the mannose 6-phosphate receptor pathway. Treatment hypothesis is to reduce or stabilize the accumulating harmful intracellular metabolites that causes NCL.
    •  Pelizaeus-Merzbacher Disease (PMD): Survive and proliferation of huCNS-SCs in vivo to produce oligodendrocytes that would generate compact myelin and to normalize nodal and axonal function.
    •  Spinal Cord Injury (SCI): huCNS-SCs transplant is proposed as a restorative therapy by providing a cell source capable of differentiating in all three respective cell lineages: astroglial; neural; and oligodendroglial.
    •  Age-related Macular Degeneration (AMD): Cell transplant to help restore cell cycle action for photoreceptor support (phagocytosis/waste processes, nutrients & neuro-protective factors) & help preserve synaptic contacts between photoreceptors, and horizontal and bipolar cells.

    Key publications:
    Direct isolation of human central nervous system stem cells. Uchida N, et al. PNAS 97:14720-14725, 2000.
    Central nervous system stem cell transplantation for children with neuronal ceroid lipofuscinosis. Selden NR, et al. J Neurosurg Pediatr. 2013 Jun;11(6):643-52
    Neural stem cell engraftment and myelination in the human brain. Gupta N, et al. Sci Transl Med. 2012 Oct 10;4(155):155ra137
    Publications are available for download at StemCells Inc. website
    Clinical Trials:
    NCL: Phase 1 USA NCT00337636 – completed, Phase 2 NCT01238315 withdrawn prior to enrollment due to “Lack of timely patient accrual”
    PMD: Phase 1 USA NCT01005004 – completed, long term follow-up NCT01391637
    SCI: Phase 1/2 Swiss/Canadian – Thoracic NCT01321333 long term follow-up NCT01725880 ; Phase 2 US A – Cervical NCT02163876
    AMD: Phase 1/2 USA  NCT01632527 long term follow-up NCT02137915
    Administration routes: Surgical procedure for access/application to brain/spine & vitrectomy to eye
    Key patents:
    huCNS-SC Methods – 6468794, 7115418, 6238922, 6368854
    huCNS-SC Composition of Matter – 5968829, 6777233, 7361505, 5851832
    (A full description & further details of the IP held is available at the StemCells Inc. website)
    IP Positioning summary:
    StemCells Inc., and it’s scientific colleagues, have had a long history in developing huCNS-SCs for cellular transplantation therapeutics and as such have a robust IP portfolio surrounding the platform. Having spent the better part of 20 years+ on the science they have established a strong position in the space as a leader. However, they are not alone in fetal based neuronal cell science, as there are a few active players in the sector with products in the clinic. The other groups in this cell source sector in the West are Neuralstem Inc. and ReNeuron Ltd., both of whom are developing CNS cell transplant products for various indications. Neuralstem and Stemcells have an ongoing legal tussle over long contested freedom to operate claims which still remains a question for both parties pending the resolution of the dispute. In regard to the eye space StemCells will soon be joined by ReNeuron/Schepens Eye Institute who also have a developing program using fetal CNS cells (hRPCs “retinal progenitors”) – however for a different indication (Retinitis Pigmentosa). In the overall CNS space there are a number of cell therapy programs in development, the most notable of which are the Pluripotent players (Ocata, BTX/CellCure/Asterias & RIKEN). These programs, and the adult cell & device products being studied in the clinic, represent market competitors rather than cell source IP competitors.

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    Cells Weekly – April 12, 2015

    by Alexey Bersenev April 12, 2015 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. Public discussions on human germ line genetic engineering From scientific discussions, calls for moratorium and journal editorials, the hottest topic – gene modification of human germ line – went public widely. Wall Street Journal’s opinion piece – Let’s Hit ‘Pause’ Before Altering Humankind, written by Nobel laureats David Baltimore and Paul […]

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    Clinical cell processing news – part 2, 2015

    by Alexey Bersenev April 9, 2015 cell product

    Clinical Cell Processing News is a series about new protocols, products and techniques for clinical-grade cell processing and manufacturing. Cell processing devices, cultureware, bioreactors, GMP-grade reagents, cell separation techniques. This series is posted every 2 months. FEATURED: Validation of Xeno-free CTS Immune Cell Serum Replacement for expansion of human T-cells (Clin Transl Immunol) FREE Dynabeads CD3/CD28 Cell Therapy System (CTS)-activated or antigen-specific T cells expanded using the xeno-free SR, CTS Immune Cell SR, showed comparable growth kinetics observed with cell […]

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    Commercialization of stem cell secretome

    by Alexey Bersenev April 7, 2015 cell product

    Stem cell secretome, collected in a form of cell culture conditioned medium (CM) or supernatant, is getting more and more attention not only from researchers, but also from industry. While growing, cells release in culture medium biologically active substances and structures, such as cytokines, growth factors, enzymes, microvesicles/ exosomes and genetic material. Therapeutic value of stem cell CM was demonstrated in many studies, reviewed here. The first clinical trials, assessing CM have been started. The results of the first clinical […]

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    Cells Weekly – April 5, 2015

    by Alexey Bersenev April 5, 2015 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. Young mitochondria maintain stemness Very interesting phenomenon was described this week in Science. During asymmetric division, stem cell segregate old mitochondria to the daughter cells, which will differentiate: Daughter cells that received fewer old mitochondria maintained stem cell traits. Inhibition of mitochondrial fission disrupted both the age-dependent subcellular localization and segregation […]

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    10 companies to watch – part 4

    by Alexey Bersenev March 30, 2015 RegenMed digest

    Welcome to the 4th issue of Ten Companies to Watch – one of the most popular series on our blog! Here I pick and overview the most interesting new companies in stem cell/ cell therapy/ regenerative medicine field. I’m focusing on startups, but considering any other “under-appreciated” companies. As always – I’m open to your suggestions. Tell me why any particular company is different and worth our attention. Ok, here is new list without particular order: 1. Unum Therapeutics Cellular […]

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    Cells Weekly – March 29, 2015

    by Alexey Bersenev March 29, 2015 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. Using CAR T-cells in solid cancer – results of clinical trial CAR T-cell therapy in oncology is the a very hot topic right now. So far, it has been very successful in treatment of hematological malignancies. However, with exception of few cases, the results of using CAR T-cell therapy in solid […]

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