• 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 […]
  • This is promotional post by Howard Choi

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    This is a guest re-blog by Michael Cea

    bob langer2 I don’t believe Dr. Robert Langer needs too much of an introduction. His work and creative mind has motivated some of the brightest scientists to reach forward and look to driving change, as an achievable goal. The Langer Lab at MIT has become synonymous with innovation and a can do attitude. A view that permeates throughout the complex interwoven fabric of chemical constructs, nanoscale architectures and biological systems. This is Bob’s culture, as he likes to be called. Those from yesterday and today all have this in common, as will those of tomorrow – the origin story of innovation without peer. The list is long and distinguished and will be so far into the future.

    The utility of material science as a foundational springboard for patient centric approaches to medical intervention is not novel. There have been devices, tools and technologies throughout the history of patient care. Some of the most important shifts in medical practice has come by way of devices. The hand of the doc always can and should always be assisted with the very best innovations science can develop.

    Up until recently that was the realm of dedicated professionals, each with their own areas of expertise. Bob was one of those that looked to change that, perhaps as a result of his requirement to improve what he saw as inefficiencies and his innate ability to bridge between the professional definitions that ordered things. To seek out compatible solutions that mixed disciplines in order arrive at a solution. The utility factor by design. Practical and purposeful from a solutions perspective.

    That is what stuck me about Bob during our meeting. His natural ease at distilling a challenge and suggesting a practical pathway to arrive at a solution. Not the only solution, but in most cases a simple solution, which interconnects various synthetic elements to arrive at a bioengineered answer. A product that can work with and within our natural biological systems, often dramatically improving on an existing methodology for the benefit of patient care.

    During our conversation we discussed two such technologies that I have been following closely: SQZ Biotech and Gecko BioMedical. In addition, I followed-up with management for further details. Below you’ll find a summary of these technologies and the associated Q&A.

    A third topic was InVivo Therapeutics, which has been extensively covered by analysts and the media so I’ll only add some color from Bob on the history and developments.

    InVivo’s origin story began with a graduate student, Erin Lavik, in the late 90s. She had come to Bob with with her thesis on spinal cord repair as a material scientist student doing her Phd. They talked about creating a scaffold that “looked like the grey and white matter of the spinal cord which we could put neuronal stem cell on.” Erin had been collaborating with Evan Snyder, a stem cell scientist and envisioned a combined biomaterial and stem cell product. The PNAS paper they published with Yang (Ted) Teng, a neurosurgeon, showed “significant improvement in rats” with the scaffold & cell product. Later they showed improvement in monkeys with both scaffold only and to a greater extent with the combined approach with cells. At that time they were approach by Frank Reynolds from MIT Sloan School about Licensing – which launched InVivo. Frank has since stepped down and Mark Perrin now heads the company.

    By Bob’s account, “I think Frank and the people at InVivo, thought it best to try to understand things properly, so they started with a scaffold only product. It’s also easier from a regulatory standpoint that way.”

    By all accounts that decision was correct, as the clinical trial has started and the first three patients have reported improvements.

    What no one expected was the first two patients “falling in love & living together – which is amazing!”

    Bob says the “long range plans are very exciting, not just for the scaffold but for the scaffold with neuronal cells and possibly the controlled release of different neurotrophic factors, which we’ve done a lot of work on, for the combined product.”

    In the meantime there is speculation not only about the source of the neuronal cells, but also about the marriage of Jesi and Jordan, those accidental love birds!

    Utility by Design, along with a touch of Karma.

    Cheers

    SQZ Biotech

    Cell in squeeze2With the advent of cellular therapy the use and in-vitro manipulation of cell populations has become a common occurrence in laboratories and bio-manufacturing centers around the world. The promise of new biologically relevant patient treatments, both personalized and generic, holds great promise for the medical industry and most of all to patients. Along with the advent of this new era in therapies comes the technologies to optimize and enhance the cell productization process.
    SQZ Biotech was born to fulfil that mission and uses it’s CellSqueeze technology to do so.

    Armon Sharei As a doctoral student in Bob Langer and Klavs Jensen’s labs in Boston, Armon Sharei excelled at the study of microfluidics, the process of passing substances through small chambers to mimic naturally occurring dynamics. During this thesis work Armon discovered that it was possible to alter the integrity of cell walls in such a manner that doorways opened into cells. Bob Langer was as surprised as Armon “when cells went through the device and a little pressure applied the cells would open up and things would go in.” After further study it was revealed that all kinds of molecules etc, large and small, would go in and no harm to the cell was observed afterwards.
    After filing patents and publishing the work in PNAS, Scientific American called it one of the world changing ideas of 2014 and the company was launched with institutional & seed support and Armon became its co-founding leader full time.

    SQZThe company has now secured VC backing to the tune of $5 million and is in industry prototype tests, with a good deal of buzz around the potential.

    Certainly the technology has got off to a good start, gaining some traction. Also there is a broad uptake in cell system technologies that can customize and safely adapt therapeutic populations. CellSqueeze could very well be the next step up in the in-vitro cell manipulation suite.

    I asked Armon a few questions on the technology -

    M: What are the best types of cells to “squeeze” and if there is a most suitable cell type model depending on the payload, i.e. primary types, differentiated cells, size or shape?
    A: We have found that the concept works with every mammalian cell type we have tried. To your point, the most promising areas have been primary cells such as immune cells and stem cells which are hard to treat by conventional means but respond very well to our technology. The size and shape of the cells do influence their delivery properties and we have a library of chip designs that accommodate different cells. As for payload, because the delivery process appears to be a largely membrane disruption based process, we can deliver a broad variety of payloads including peptides, proteins, polymers, DNA, RNA, nanoparticles, etc.

    M: How would you best describe the squeeze technology and can it be considered a transduction system?
    A: We have tended to simply call it a “delivery system” or an “intracellular delivery system”. Mostly because transduction tends to be used in the context of viruses and transfection implies nucleic acid delivery.

    M: I’ve noticed a few new systems for delivering payloads into cells – what advantage do you see SQZ having over these other developing technologies? e.g. the UCLA Optofluidics system is looking to do something similar & how do you see these different approaches vis-a-vis SQZ…
    A: I think we are going through a period of exciting development in MEMS technologies for intracellular delivery. I have seen reports of many exciting new methods such as nanoneedles, microfluidic electroporation and other physical delivery systems. Ultimately I am most optimistic about our technology because it is robust, simple, and scalable by comparison to other approaches. For example, the nanoneedles can be difficult to fabricate and are not well suited for suspension cells while microfluidic electroporation does not overcome some of the inherent toxicity issues and delivery limitations of conventional electroporation. In contrast, our devices can operate at over 1,000,000 cells/s and have demonstrated applicability to over 25 cell types. Moreover, our papers demonstrate 10-100x greater performance compared to conventional approaches in multiple applications.

    M: It has been reported that SQZ is actively engaged in trialing its proprietary system with industry – can you expand a little on that?
    A: We have active partnerships with multiple pharmaceutical and biotech companies pursuing applications that are uniquely enabled by SQZ’s technology. These often involve delivery of materials that cannot be introduced into cells by conventional means.

    M: With your recent capital raise what are your next development steps and timelines?
    A: We are accelerating our internal therapeutic programs and developing next generation devices. The company is focused on the development of novel cell therapies using multiple cell/molecular engineering modalities to address acute clinical challenges across indications.

    Refs:

    Gecko Biomedical

    As the name inspires, Gecko is a product of nature, developed as a mechanistic solution to the closure and repair of tissue in wet conditions.

    As I recall, I was drawn to this technology as a result of my father’s experience as a surgeon and his constant struggle with closure and repair. From his days as a field surgeon in Vietnam to his surgery days at the hospital, he always wished there were more effective closure tools so he could perform better. Occasionally I would listen to him speak about those young soldiers.

    The need is real and the solution a potential breakthrough. To be able to close an internal or external wound with the ease and elegance of a bio-patch is a proactive step forward.

    Jeff Karp Langer again was instrumental in providing the fertile ground, this time for post-doc Jeff Karp, now an Associate Professor at Brigham and Women’s Hospital, Harvard Medical School. As Langer relayed, Jeff loved “to take things out of nature and make them into synthetic materials and one day we were talking about Geckos and if we could make a polymer similar to a Gecko’s grip and we did by nano-printing.” That was the start of development and since then the technology has evolved with advanced glue inspired by snails & worms, along with the integration of light activation chemistry.

    Gecko Biomedical is based in Paris and concluded a successful private placement with VCs for $11m, and is running preclinical studies in preparation for first in human trials later this year. Another Langer venture, Moderna, a mRNA company, was the catalyst for the French connection, as he was introduced to Gecko’s future management via Moderna’s executive suite.

    I asked Jeff Karp for a little background on the technology and its applications.

    M: Can you explain a little about the product solution
    J: There is a huge unmet need for better tissue adhesives. Sutures are extremely time consuming as with each pass of the suture needle, the tissue needs to be re-aligned. And the longer a patient is on the operating table the greater the chance for complications. It is also difficult to tie knots in small spaces, such as during laparoscopic procedures. Staples are also problematic as anytime one pushes a staple into tissue, the hole that is created is larger than the staple which tears the tissue, and this can serve as a nidus for bacterial infiltration. And typically you need to bend staples to secure them in place which damages the tissue. Often staple tracks are sites of infection! Also it is challenging to apply staples in small spaces as the devices used can be quite bulky. And sutures and staples have different properties than tissue and this mismatch can cause tissue death over time and lead to leaks.

    M: How did you develop the design requirement for the platform
    J: We put together a design criteria for the solution -
    Gecko img

    • We wanted this to work in the harshest environment inside the body, inside a beating heart; it is a highly dynamic environment, it’s very wet, lots of proteins. Many of the glues that exist today that are being used for tissues can become fouled in the presence of blood. As soon as they contact blood they can no longer adhere to tissue so we had to solve that problem.
    •  We wanted it to be degradable and Biocompatible. What I mean by this is because we are trying to treat kids, we wanted this to facilitate the migration of cells overtop and into it so as the material degrades, it would be replaced with the patient’s own tissue. So at the end of the day, maybe 4 months or 6 months, you’d just be left with the patient’s tissue and that can grow overtime so you don’t have to come in and do revision surgeries.
    • It needed to be elastic as the heart is undergoing multiple expansion/contraction cycles.
    • Also, we wanted this to resist washout in the heart which is very challenging because there is a high shear stress.
    • When we talked to a number of clinicians, they said “There’s certain glues that exist in the clinic that either cure within 1 minute or 10 minutes and we don’t want to be at the mercy of the technology, we want to be in control”; and so we made on-demand adhesion part of our design criteria. We envisioned using light to achieve this.
    • We had materials that could address many of these, but not washout & adhesion. We couldn’t figure out a way around it so we turned to nature for inspiration and we synthesized glue inspired by slugs snail and sand castle worms. We produced a viscous prepolymer, that you would apply to the tissue or to a patch and then cure with light to end up with a material that’s very similar to an elastic band. We made this patch you can stretch it over and over again but it’s fully degradable and biocompatible. It can take a 30% strain, which is what you’d expect in the heart, and we don’t see much change in the mechanical properties of the material. Eventually we showed that we could seal the carotid artery and aorta of a pig, and also attach a patch inside a beating pig heart.

    c bancelI also asked Christophe Bancel, CEO of Gecko Biomedical, a few questions on the formation of Gecko and the technology platform moving forward.

    M: How did you know of the technology
    C: Bernard Gilly (Non-Executive Chairman) and I knew Bob and Jeff Karp and their the work in the field of adhesives in wet and complex environment for a certain time and we followed their progress. Once the 4 of us believed that the technology was ready to start translation into a product, we decided to create Gecko Biomedical.

    M: Why Paris and not the US?
    C: From a regulatory and development point of view, many innovations in Medtech tend to be developed in Europe first, so we thought that this could be an opportunity. On top of that, Bernard had developed an entrepreneur initiative in Paris, the iBionext Network, that had successfully brought together experienced executives in Biotech and Medtech with support of leading European investors. We were ready to provide the full support for the development of this technology into innovative products for patients.

    M: How do you view the underlying technology
    C: Gecko Biomedical’ platform allows the development of diverse solutions for adhesion and wound closure in wet and complex environment that can be designed to meet the requirement of specific tissues. Applications range in the different fields of surgery. We have decided to start focusing on vascular reconstruction, but are also developing variants of our polymers for new tissue types.

    M: Will there a combined product with biologics in a future generation of products?
    C: Indeed, by design, our family of polymers can provide controlled release of active substances (small molecules or biologics) and also encapsulate cells for active delivery.

    M: Where are you at present in the product development timeline?
    C: Currently we are working with a leading cardiovascular department at Paris Hospital. We are finalising all the regulatory development under Good Laboratory Practice (GLP) for the non-clinical validation of our first application in vascular reconstruction and intend to start the first clinical trial by year end.

    Refs:

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    Cells Weekly – July 26, 2015

    by Alexey Bersenev on July 26, 2015 · 1 comment

    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!

    CD34+Banner_490x90

    1. New synthetic feeder-mimetic induces robust expansion of stem cells
    A group of researchers from U of Nottingham (UK) discovered and synthesized new polymer, which induces expansion of human pluripotent stem cells. They used high throughput screening for library of polymers to identify the best candidates to support human ES cell line in StemPro media. They ended up with co-polymerization of two candidates. The beauty of the method is in its robustness – expansion of stem cells was independent of culture labware, media, preconditioning (ECM protein preadsortion) and type of cell line (ES/ iPS):

    Thus, poly(HPhMA-co-HEMA) fulfills all the current culture requirements for the clinical use of stem cells within regenerative medicine and can be scaled up by coating onto cultureware to be used off-the-shelf, providing a cost-effective alternative to commercially available hPSC expansion substrates.

    2. Introducing cryobioprinting
    Researchers from U of Manchester, for the first time, bioprinted cells, mixed with cryoprotectant DMSO for immediate cryopreservation of printed construct. They named their new method cryoprinting. They use 3 different methods: (1) printing cells on substrate at room temperature with following freezing at -80C, (2) printing cells on pre-chilled to -80C substrate (on dry ice) and (3) direct cell printing into liquid nitrogen:

    After printing and a freeze/thaw cycle (with a minimum 24 hours hold period at liquid N2 temperature), 3T3 cells showed an average viability of >90% with CPA concentration

    Cell viability was not as good when they used neural cell line and method 3.

    3. Educational online course – Stem Cell Therapeutics
    For the first time, famed academic institution – Stanford University, offers an online education course on stem cells. You can see announcement in this video. By the end of the course Stanford Genetics and Genomics Certificate will be provided. A price is $495. If you will enroll, please share your experience with a course!

    4. Update on Jacob Hanna’s papers issues
    Retraction Watch gives an update this week on multiple issues, related to published papers of prominent Israeli stem cell scientist Jacob Hanna:

    Last month, commenters on PubPeer noticed that images from at least 10 of the research papers Hanna coauthored in seven journals — that commenters had posted on the image hosting website Imgur and linked to on PubPeer — had been deleted.

    5. Investors to sue gene therapy company
    Public gene therapy company Avalanche Biotechnologies announced mixed unimpressive results of their Phase 2 clinical trial in June. This week, Avalanche’s CEO resigned. Also this week, Avalanche’s investors started an investigation:

    The investigation focuses on whether the Company and its officers violated securities laws by misleading investors regarding the strength of its Phase 2 clinical trials, and hid the fact that the trial was not designed to show statistically significant differences between the control and active groups.

    6. How CAR T-cell technology can save stem cell company
    Very interesting story about French biotech company Cellectis was recently posted on LabioTech web-site. CRISPR came and knock their business down, but then CAR came:

    A much cheaper alternative to Cellectis’ meganuclease that has to be custom-made for each customer. The emergence of its new rival led Cellectis’ revenues to sink down to alarming rates – the French company reported a net loss of €61,7M for the year 2013 alone. The company had no other choice, but to announce a massive restructuration plan and to close down its kit/bioresearch activity, abandoning over 100 employees and refocusing on therapeutics in the oncology space.
    A wise move looking back at it now, but then again, it was pretty risky.
    The company was 3-months away from bankruptcy, had little data in pre-clinical and nothing in humans. But its expertise in gene editing could be transposed into the cancer field – in the trending field of CAR-T…

    A few stem cell companies slipped to CART and other immunotherapies. Have you noticed this trend?

    7. New methods and protocols:
    Prospective isolation of human erythroid progenitors (PNAS)
    Single-step generation of conditional knockout mouse ES cells (Cell Reports)
    High-throughput screening of surface marker human adipose-derived stromal cells (Tiss Eng Part A)
    A pre-screening method to detect CRISPR/Cas9 off-targets in mouse ES cells (Sci Reports)

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    Everyone who is entering the field of clinical expansion of mesenchymal stromal cells (MSC) facing the problem: available reliable supply of serum for cell culture. As number of MSC clinical trials is increasing with years, the global demand for serum is growing dramatically. Taking into account prevalence of using serum for clinical MSC culture worldwide and “peak serum phenomenon”, it becomes clear that this “serum model” is not going to work in the future. There will be not enough supplies of clinical grade serum (animal or human), serum-based MSC manufacturing model will be unsustainable and, eventually, obsolete. It is also clear to me that serum alternatives or completely synthetic media should be tested early during product development. New serum-free formulations may have advantages over serum or alternatives, but not many products available to test, since industry in its early days. That’s why every good study on this topic is a worth our attention! Today I’ve picked one of such studies, recently published in Cytotherapy.

    The authors tested hypothesis of finding one “ideal” serum-free media (SFM) for different MSC cultures. They tested 6 SFM on 7 different MSC lines in 3 different culture methods. Tested SFM:

    • StemPro MSC SFM Xeno-free (Life Technologies/ Thermo)
    • MSC Nutristem XF (Biological Industries)
    • MesenCult-XF (StemCell Technologies)
    • StemXVivo SFM (R&D Systems)
    • 2 in-house SFM fromulations (Biotechnology Processing Institute)

    Human MSC cell lines were obtained from:

    • Promocell
    • ATCC
    • Medipost
    • CTI Biotech
    • RoosterBio
    • NUH,Singapore

    Tissue sources of these lines were:

    • umbilical cord matrix
    • umbilical cord blood
    • adult bone marrow
    • fetal bone marrow
    • adipose tissue

    Cell culture methods:

    • monolayer
    • 3D static microcarrier
    • 3D agitated microcarrier

    Now, I’d like to summarize findings from this study:

    1. In monolayer culture MSC density/ proliferation did not correlate between SFM and serum+.
    2. In terms of MSC density, most SFM outperformed serum+.
    3. There was no single SFM optimal for growth of all tested MSC lines.
    4. Cell density/ expansion differ significantly between different SFM.
    5. The optimal SFM differ between monolayer and microcarrier-based cultures.
    6. There was no correlation in MSC growth between static and agitated microcarrier cultures.
    7. There was a poor correlation between MSC growth in monolayer and microcarrier serum+ culture.
    8. Some of MSC lines were slowly growing and none of SFM supported them on static microcarriers.
    9. Cell attachment onto microcarriers may not predict good proliferation in SFM.

    Take home messages from this study:
    Optimize SFM for your particular MSC type, method of expansion, other culture supplements, coating matrices and application! I couldn’t emphasize this more! There is nothing more important for MSC developer than cell culture optimization! Find your own “magic formula”! It worth your time and money! Do it early on – before even starting Phase 1 of trial. Try all available options – commercial/ in-house-made SFM formulations and serum alternatives. Use lower (1-2%) and high (5-20%) concentration of serum as controls. Don’t rely on publications, don’t make assumptions – test it on your own! Your adipose/ marrow MSC could grow very differently from other reported adipose/marrow MSC in the same culture conditions.

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    Cells Weekly – July 19, 2015

    by Alexey Bersenev July 20, 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. Fetal tissues trading scandal Whole week, US mass media covered a scandal, related to aborted fetal tissues trading by planned parenthood. At the heart of the scandal is a leaked video, recorded by anti-abortion group of activists, where planned parenthood’s senior director of medical research discusses details of harvesting of aborted […]

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    Niche conditioning for stem cell engraftment

    by Alexey Bersenev July 16, 2015 methods

    As you may know from this blog, host conditioning is required for successful engraftment of hematopoietic stem cells in BMT models. Conditioning allows to prep bone marrow niche for donor stem cells by depleting host stem/ progenitor cell pool. A typical niche conditioning in BMT models (as well as in clinic) is done by total body irradiation (TBI). Besides TBI, antibody- and chemotherapy- based conditioning is also used in experimental models and in the clinic. Successful niche conditioning is a […]

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    Cells Weekly – July 12, 2015

    by Alexey Bersenev July 13, 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. Stem cells self-made niche for progeny It is known that stem cells can make niche cells for themselves. A new study revealed reverse relationships between parent stem cells and daughter progeny: Using a combination of cell ablation, lineage tracing and signalling pathway modulation, we show that airway basal stem/progenitor cells continuously […]

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    Divesting of VSEL

    by Alexey Bersenev July 9, 2015 other adult stem cells

    About a month ago, US-based NeoStem, which was trying to commercialize VSEL technology, was re-branded to Caladrius. With rebranding, however, information about VSEL development platform was removed from Caladrius web site. There was no any official statements from the company about VSEL divestment, but apparently they did it. I’m curious to learn about reasons for VSEL divestment, but I can only guess: Did Weissman’s paper play role and discredit VSEL? Stuck in translation? Were any reproducibility issues? Did company decide […]

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    Video interview with Jennifer Doundna on discovery of CRISPR-based genome editing

    by Alexey Bersenev July 9, 2015 educational

    Jennifer Doundna is a person behind “biotechnology discovery of 21st century” – CRISPR-based genome editing. Besides great science, she is highly involved in public debate on current state of CRISPR technology and ethical issues, related to genome editing of human germline (see her testimony for US Congress). Finally, she is involved in commercialization battle of CRISPR-based technologies. This video was recorded on the recent 2015 Cold Spring Harbor Symposium “21st Century Genetics”. I especially like the story behind of discovery […]

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    Cells Weekly – July 5, 2015

    by Alexey Bersenev July 6, 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. CRISPR genome editing news Famous geneticist from MIT Eric Lander wrote an opinion piece in NEJM. I’d highly recommend you to read this article! The discussions that will begin in the fall may solidify a broad international consensus that germline editing should be banned — with the possible exception of correcting […]

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