• 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 […]
  • Clinical cell processing news – part 6, 2014

    by Alexey Bersenev on December 17, 2014 · 0 comments

    in clinical lab

    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.


    Validation of “G-Rex M series” system for expansion of suspension cells (Molecular Therapy Methods) FREE

    A multicenter study confirmed that this fully optimized cell culture system can reliably produce a 100-fold cell expansion in only 10 days using 1L of medium. The G-Rex M series is linearly scalable and adaptable as a closed system, allowing an easy translation of preclinical protocols into the good manufacturing practice.

    1. Validation of 96-hours room temperature storage of cord blood before processing (Vox Sanguinis)

    The delay of 96 h before UCB processing is possible, without a prohibitive impairment of CD34+ loss in number and functionality.

    2. Use of autologous plasma for cryopreservation of hematopoietic cells (Transplant Proc)

    The results show that the application of autologous plasma is linked with higher viability of nuclear cells and CD34+ cells. Moreover, statistical analysis of the nuclear cells and CD34+ cells viability differs significantly between groups frozen using RPMI 1640 and autologous plasma (P < .05).

    3. Validation of improved cord blood processing with Sepax system to yield increased number of cells (Cytotherapy) FREE

    This report addresses the challenge of recovering consistently high TNC yields in volume reduction by proposing and validating an alternative protocol capable of processing a larger range of units more reliably.

    4. Development of potency assay for immunomodulatory qualities of MSC (Cytotherapy)

    We measured MSC-induced suppression of CD4+ T-cell proliferation at various effector-to-target cell ratios with the use of defined peripheral blood mononuclear cells and in parallel compared with a reference standard MSC product. We calculated an IPA value for suppression of CD4+ T cells for each MSC product.

    5. Development of immunosuppression potency assay for cell products (ASH 2014 abstract #316)

    We developed an in vitro standardized quantitative suppression assay utilizing the suppressor cell line KARPAS-299 as a standard by which to compare suppressive potency of human immune-regulatory cell populations.

    6. Relocation of cord blood bank units using high-capacity dry shipper (Transfusion)

    In this report, we share our experience on relocating more than 100,000 units of cryopreserved UCB samples stored in 12 liquid nitrogen freezers (LNFs) to our new laboratory.

    7. Increase TNC yield from cord blood by adding a Hespan (ASH 2014 abstract #1126)

    AXP-automated CB processing with HES addition can be performed in the GMP environment, results in higher post-processing TNC and therefore, increases the CB bank’s ability to store larger CBUs that are most useful to patients.

    8. Validation of sterility control for cellular products according to German regulation (Vox Sanguinis)

    In order to generate standardized conditions for the microbiological control of HPCs, the PEI recommended defined steps for validation that will lead to extensive validation as shown in this study, where a possible validation principle for the microbiological control of allogeneic SCPs is presented.

    9. Quality control system for cell products in China (Tissue Engineering Part A)

    In this review, we first briefly introduced the major institutions involved in regulation of cell substrates and therapeutic cell products in China, and the existing regulatory documents and technical guidelines used as critical references for developing the new interim guidelines. With focus only on non-hematopoietic stem cells, we then discussed principal quality attributes of SCMPs as well as our thinking of proper testing approaches to be established to ensure all quality requirements of SCMPs along manufacturing processes and development stages.


    Cells Weekly – December 14, 2014

    by Alexey Bersenev on December 14, 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!


    We are planning few posts about results and achievements of the year. Please send your suggestions about “most significant events of the 2014″ in stem cell research and regenerative medicine. As the first step, I’d like to ask you to take a survey “The best method of 2014“. You can vote here.

    1. F-class of pluripotent stem cells described
    Number one news in stem cell world this week was a discovery of new type of pluripotent stem cells – so called “F-class”. The major study was published in Nature and 3 more in Nature Communications. All of them are parts of one big project, led by Canadian scientist Andras Nagy. You can look at raw data here.
    These studies were covered very nicely, I’d suggest you to read Nature News, the Scientist and Knoepfler Lab blog.

    2. More insight into Macchiarini’s case investigation
    If you follow the allegations case of Paolo Macchiarini in research misconduct, you may be interested in reading documents, obtained this week by Retraction Watch:

    As supporting evidence, the four doctors also produced an analysis of six of Macchiarini’s papers. Each one lists inconsistencies and inaccuracies, including:
    1. Inquires have been unable to identify any application for synthetic tracheal transplantation filed at the Regional Ethical Review Board.
    2. Unapproved informed consent form signed 17 days after the transplantation.
    3. Serial fabrication and omission of biopsy findings.
    4. Serial fabrication and omission of bronchoscopic findings.
    5. Fabrication and omission of clinical status.

    3. Summary of ASH 2014
    This week, the biggest annual hematology conference – ASH, finished in San Francisco. I was following it via twitter and was pleasantly surprised by number and quality of real time tweets and blogging. There were a lot of important and interesting clinical reports, mostly in cellular immunotherapy of cancer. Here I pick the most interesting cell therapy-related posts about ASH 2014:
    Cell therapy highlights from #ASH14 (CellTrials.info)
    CAR T-cell tweets from #ASH14
    Immune cells boost cancer survival from months to years (Nature News)
    ASH – Novartis, Juno, June and Rosenberg steal the T-cell show (EP Vantage)
    Immunotherapy Confidence (And Audience) Abundant At ASH T-Cell Talk (Xconomy)
    Bluebird Gene Therapy Inducing Durable Cures in Blood Disorder Patients (the Street)
    CAR T-Cell Therapy Elicits High Response Rate in Pediatric ALL (CURE magazine)
    Hematopoietic clone tracking may help guide therapies for malignancies (HemOnc Today)
    ASH Roundup: Immunotherapy Stars, Gene Therapy Leaps Ahead (Xconomy)

    4. CRISPR patents battle
    The first candidate for the “Method of the Year 2014″, genome editing technique CRISPR, is undergoing very interesting historic moment – a battle for intellectual property. MIT Tech Review posted very interesting piece:

    How did the high-profile prize for CRISPR and the patent on it end up in different hands? That’s a question now at the center of a seething debate over who invented what, and when, that involves three heavily financed startup companies, a half-dozen universities, and thousands of pages of legal documents.

    Also this week, Nature Biotechnology posted an article – Who owns CRISPR-Cas9 in Europe?

    The Zhang patent has recently been granted in the United States. In Europe, it will take a long time before the extent of the patent monopoly owned by the Broad and MIT is clear. This also applies to the other existing CRISPR-Cas9 patent applications. Moreover, as is illustrated here, the claims of the Zhang patent and others may very well differ between the United States and Europe. All of this adds to the increasing uncertainty surrounding the CRISPR-Cas9 patent in Europe. Third parties in need of access to this scientifically promising technology should be aware and should carefully assess their legal and commercial options.


    5. Self-assembled biomaterial modulates immune cells in vivo
    David Mooney’s lab created injectable self-assembling biomaterial, which able to modulate immune response in vivo. The study is published this week. This is very interesting concept:

    … we propose an approach in which host immune cells are recruited and modulated in vivo by 3D scaffolds that spontaneously assemble from mesoporous silica rods (MSRs) of high aspect ratio…
    … we describe injectable pore-forming scaffolds based on MSRs, and demonstrate their application to in vivo modulation of host immune cells and potential as a vaccine platform to provoke adaptive immune responses.

    6. Cancer stem cells for beginners – markers and flow cytometry
    Great explanation of cancer stem cell surface markers was posted on the Signals blog:

    Sample preparation can cut off surface markers of interest. Cancer cells are ‘sticky,’ so they need to be separated into single cells before going through the flow cytometer (single file, remember!). Sometimes the techniques used to do this can cut off surface proteins – including the marker of interest – so the cell may falsely appear negative for the marker.

    Highly recommended to all beginners!

    7. Retraction of the week
    Retraction Watch reports about retraction of “stem cell paper” from Korean journal, linked to scandalous “Stamina case”:

    A Korean stem cell journal has retracted a paper on a controversial Italian treatment that involves harvesting stem cells from bone marrow and injecting them back into the patient.


    The most interesting methods of 2014

    by Alexey Bersenev on December 13, 2014 · 0 comments

    in methods

    Every year we pick the best methods in stem cell research/ regenerative medicine. I’d like to ask – what methods were the most interesting/ significant this year for you? Please suggest the methods which are relatively new, but not necessarily described this year. I’d like to hear about the methods, which got some “kick” or advanced significantly (moved to clinic or business). Here is a survey, where I included few methods, which I’ve mentioned on a blog in 2014. Please take a survey or suggest some other methods in comments! I’m going to pick “the best method” in about 2 weeks. Stay tuned!

    CLARITY-like techniques
    Molecular beacons
    Barcoding in situ
    CRISPR genome editing


    Products in the pipeline – Cell Cure’s OpRegen

    by Alexey Bersenev on December 10, 2014 · 1 comment

    in cell product

    This is third 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
    (updated: 12/15/2014)

    Product candidate name: OpRegen®
    Developer: Cell Cure Neurosciences Ltd. of Israel (a subsidiary of BioTime, Inc. with Teva Pharmaceutical Industries, Ltd. and HBL-Hadasit Bio-Holdings, Ltd. as minority shareholder partners)
    History of development: Since 2005 by Cell Cure Neurosciences Ltd. (a ES Cell International Pte Ltd) acquired by BioTime, Inc. in 2010
    Type of cells: Human allogeneic embryonic stem cell-derived retinal pigment epithelium (RPE), expanded ex vivo.
    Tissue source: Inner cell mass of blastocyst, derived from fertilized oocytes. Pre-embryo donated as supernumary tissue to science rather than discarding during standard IVF treatment. hESC cell line established – HAD-C 102.
    Processing steps: Laser blastocyst ZP breaching, mechanical dissection, isolated ICM plated, ES culture to embryoid body development on human foreskin fibroblasts feeder layers in KO medium, ES cells isloation, hESC master cell bank, further culturing steps w/ nicotinamide and TGF-β Growth Factors (Activin A) until RPE cells, plating, expansion, working cell bank, expansion, harvest, cryopreservation.
    Phenotypic composition: positive for MiTF-A, RPE65, Bestrophin, Mertk, Otx2, ZO-1, CD81 and CRALBP
    Stability/Safety: Genetically stable, normal karyotype, normal senescence & terminally differentiated to its RPE fate
    Proposed mechanisms of action: Cell replacement to restore cell cycle action to support photoreceptor.
    Scientific background: RPE cell replacement therapy is a potential regenerative treatment which looks to restore lost retina cells at the base of the macula to establish a new and healthy support layer to the photoreceptors which are loosing function as a result of the old RPE layer dying off. It is envisioned that by replacing the lost RPE layer the remaining photoreceptors will once again function properly thereby either arresting the decline of the age related condition or actually improve lost visual acuity.
    Key publications:
    PLoS One. 2012;7(6):e35325. doi: 10.1371/journal.pone.0035325
    Cell Stem Cell. 2009 Oct 2;5(4):396-408. doi: 10.1016/j.stem.2009.07.002
    Ongoing trials: Phase 1 – Dry AMD – Geographic Atrophy – NCT02286089
    Administration: Vitrectomy with sub-retinal injection of cell suspension with dosing from 50,000 to 500,000 cells
    Key patents:
    hESC Culture System – WO2008120218 & US Patent 8,597,947 & US Patent App 13/357948 (Notice of Allowance issued)
    Stem Cell Derived RPE Cells – WO2008129554 & US Patent App 12/450943 & Espacenet Family
    Method of Selecting RPE cells – WO2013114360
    IP Positioning summary: Cell Cure’s underlying US Patent on Pluripotent Culture System methods (hESCs from ICM Blastocyst stage) employs a propriatary xeno-free feeder system and is therefore a valuable addition to the state-of-the-art science to maintain Pluripotent embryonic cells for further derivation into somatic cell lineages. Biotime has a leading position in hESC Blastocyst IP as a result of Geron’s IP assets in hESC science which Biotime acquired. In addition it should be noted that the process of extracting hESCs from Blastocyst ICM stage embryos results in their termination – a point that restricts the IP due to European objection to destructive methods of embryonic cell extraction.
    The RPE derivation methods defined by Cell Cure have been issued Patents in various International Territories but are not, as of writing, protected in the US due to various USPTO Examination issues, including prior disclosure of the use of Nicotinamide in the above cell culture patent literature and Ocata’s lead in issued patents & prior art in RPE IP.
    In relation to the sector competition please see the following link for an overview: BTX Hadasit.

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    Cells Weekly – December 7, 2014

    by Alexey Bersenev December 7, 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. World Stem Cell Summit overview This week, annual meeting World Stem Cell Summit took place in San Antonio, Texas. You can read all tweets from the conference via #WSCS14. Unfortunately, this year, there were not many interesting blog posts, but I’d encourage you to read a few on CIRM’s “The Stem […]

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    Commercialization of organ decellularization

    by Alexey Bersenev December 6, 2014 RegenMed digest

    Organ and tissue decellularization (decell) is a process of cell removal with an aim to provide natural extracellular matrix ECM) for transplantation. The method is not new, but it gained momentum in the last few years with introduction of new decell techniques for perfusion of whole vital organs, such as heart, lungs and liver. Notably, in the last 5 years, the first startups, aiming to commercialize decell tissue and organs, came up. Today I’m going to overview a few companies, […]

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    What is the best reprogramming method?

    by Alexey Bersenev December 2, 2014 embryonic/iPS

    Since the first report of iPS generation technique by Yamanaka in 2006, there were plenty of other methods described in the literature. So-called “non-integrating techniques” are more attractive in terms of clinical translation. A new study, published yesterday in Nature Biotechnology, provides a great guideline for choosing the most suitable (to you) non-integrating reprogramming method. If you need to set iPS cell generation technique in the lab and have no idea what method to choose – this paper for you! […]

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    Cells Weekly – November 30, 2014

    by Alexey Bersenev December 1, 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. Paolo Macchiarini is accused of misconduct It happened again. Famous surgeon and tissue engineering pioneer Paolo Macchiarini, got into trouble. In 2012 he was accused in attempt of fraud and was arrested. This week, New York Times broke a news about possible involvement of Paolo Macchiarini in research misconduct. The Karolinska […]

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    Cell manufacturing as a generator of new jobs

    by Alexey Bersenev November 28, 2014 RegenMed digest

    RegenMed Industry Digest sponsored by: It’s an imperative in any successful economy to develop and maintain strong domestic manufacturing industries. Because of economic crisis of 2008, US, as many other countries, are “hungry” for new secure jobs in manufacturing. I believe that cell therapy and regenerative medicine, as new thriving pillar of healthcare, is providing a great opportunity for generation of many manufacturing jobs. Down the road it could boost country’s economy. Here, I’ll try to give you few examples […]

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    Lecture: Masayo Takahashi – First application of iPS cells in human

    by Alexey Bersenev November 27, 2014 lectures

    Today I was enjoying watching Masayo Takahashi’s talk, recorded on November 26, 2014. Masayo Takahashi is a scientist and physician behind the world’s first transplant of iPS cell-derived product (Retinal Pigment Epithelium – RPE) in human. A teaser – by watching this video you may learn: Why RPE cells is an ideal product for the first even iPS cell application in human How long does it take to manufacture clinical iPS-RPE product for transplant What was Sasai’s role in moving […]

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