March 2019

 
 
 

A special thanks to this issue's contributors: Dr. Peter Kilbride, Dr. Krishnaa Mahbubani, Dr. Bradley Weegman,

Dr. Alireza Abazari, Dr. Simona Baicu, Mitch Rostad, and Dr. Kate Franz

Edited by Dr. Alyssa Ward

Email alyssa.ward@organpreservationalliance.org with news, comments, or questions about the Briefings.

Click here to see the past Biopreservation Briefings!

 
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The Biopreservation Core Resource is hosting a short course on the preservation of cellular therapies. The course focuses on topics ranging from protocol development and facility design to regulatory issues in cell preservation. Click  here  to find out more information and to register.

The Biopreservation Core Resource is hosting a short course on the preservation of cellular therapies. The course focuses on topics ranging from protocol development and facility design to regulatory issues in cell preservation. Click here to find out more information and to register.

 

ASME lAUNCHES NEW E-LEARNING COURSE: CELL MANUFACTURING FOR ENGINEERS

The exponential growth of the cell therapy industry has resulted in a demand for the solutions to improve scalability, automate processes, reduce labor costs, and maintain quality assurance, which can be solved by applying engineering principles.  

ASME is pleased to introduce a new online course, Cell Manufacturing for Engineers, addressing the critical need to integrate biological concepts and engineering processes by. It is designed specifically to provide an education on cell culture techniques, manufacturing and production processes, and regulatory and other business requirements.

Engineers, bioengineers and businesses can part of the revolution of healthcare by delivering solutions toward automation, reproduction and scalability of cell manufacturing.

Please visit us at https://aabme.asme.org/learning for course availability and to sign up for updates. Your free subscription includes access to Frost & Sullivan market insights as well as exclusive content on emerging topics and the latest innovations.

 

ISBER and OPA are happy to introduce the Living Biobank Special Interest Group

Following the success of our roundtable discussion at the annual meeting of the International Society for Biological and Environmental Repositories (ISBER), OPA and ISBER have partnered to launch a Living Biobank Special Interest Group. This coalition of biobanking experts and advocates will explore the applications for biobanking created by cryopreservation of living, functional brain tissue, tumor samples, organ slices, primary cells, and other biological systems. This program will aim to outline how to implement these advances to transform key aspects of biomarker discovery and validation, personalized medicine, drug toxicity testing, and functional studies of diseased vs. healthy donor tissue.   

As the research into scaling cryopreservation to bank larger tissues and organs continues to develop and expand, biobanks may be some of earliest end-users of these technologies, due to relatively modest technical barriers. We expect to serve as a platform to facilitate collaboration among ISBER members and other stakeholders invested in viable cryopreservation. If you are excited by the opportunity to shape this Special Interest Group or to learn more about it, please contact: kate.franz@organpreservationalliance.org.

 

American Society of Transplantation webinar on ultra-rapid rewarming

In a newly released webinar, Dr. John Bischof discusses his solution to a longstanding problem in cryopreservation: how to quickly rewarm a tissue that was preserved without ice (spoiler: he uses metal to transfer heat so fast that ice can't form).

The webinar is part of the "Needs and Leads" webinar series organized by the American Society of Transplantation (AST)'s Recovery and Preservation Community of Practice (RAP COP). RAP COP was launched last year as a partnership between AST and OPA in response to the White House Organ Summit.

To join the community and view this and other webinars, click here. Or click here to access the community hub if you're already an AST RAP COP member. To nominate a topic or speaker (including yourself), email mitch.rostad@organpreservationalliance.org.

 

American Society of Transplantation (AST) Webinar on providing organs and tissues for research

Recently, leadership at the National Disease Research Interchange gave a presentation for the AST Recovery and Preservation Community of Practice (RAP COP) community on their exciting and impactful work that gives non-transplantable organs a second chance to save lives by matching them with significant medical research. 

Their presentation can be found in the AST RAP COP "Needs and Leads" webinar library. RAP COP was launched last year as a partnership between AST and OPA in response to the White House Organ Summit.

Our sincerest thanks to: 

Dr. Bill Leinweber - President & CEO 

Dr. Thomas Bell, MS, PhD - Vice President, Operations and Director, Biorepository

Dr. Gene Kopen, PhD - Senior Vice President,Strategic Initiatives

Ms. Eileen Falchetta - Director of Marketing and Communication

For access: click here to join AST and the RAP COP or here if you're already an AST RAP COP member (RAP COP library tab on community hub).

If you want to suggest a topic or speaker (or present one yourself) email mitch.rostad@organpreservationalliance.org.

 
 
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Jump to: OPA updates or News from our network

Recent papers that caught the eye of our multi-institute Organ Banking Journal Club. If you're interested in joining, contact alyssa.ward@organpreservationalliance.org to learn more!

 

A KNOCKOUT FOR CPA TOXICITY

Cryoprotectant agents (CPA) are crucial to inhibit ice formation and enable viable storage and vitrification of cells and tissues at cryogenic temperatures. Vitrification in particular requires very high concentrations of CPAs, but their application is limited by the toxicity of the compounds– increasing concentrations are more viscous and take increasing amounts of time to load and more time causes decreased viability and toxic effects. Solving CPA toxicity would not only eliminate the viability problem during loading but would catalyze solutions to other existing barriers to widespread organ banking, for example freeze-injury and ice formation.

If cellular pathways that protect during CPA exposure were identified, this could allow for either pharmacological intervention during CPA loading or genetic modification of bioengineered organs to enhance resistance and ultimately help bank organs for transplant. In a new study by Cypser et al., they used a genetic approach to identify such resistance pathways. Using transposon-mutagenesis of mouse embryonic stem cells and selecting colonies in M22, a CPA used in vitrification, they identified 9 possible mutations that significantly increased viability during CPA exposure. Interestingly, they found that some of the mutations were broadly protective against multiple CPAs and mutated cells were more resistant to freeze-thaw cycles, suggesting that the mutations protected against freeze-injury in addition to toxicity.

Although none of the hits suggested a specific or unique protective pathway, some hits like Hes1 and Myc have known pharmacological modulators that could be further tested for a viability benefit.

 

Perfusion Continues to Beat Cold (Static Storage)

Human tissue can be damaged if it lacks oxygen and nutrients, and the duration of damage determines whether the tissue will survive or die. This damage occurs during tissue, or graft, transplantation – e.g. once the tissue is removed from the body’s blood supply that provides nutrients and oxygen, a clock is running, and damage is accumulating. Grafts are therefore preserved throughout the transplantation process to slow down the accumulation of damage. Some grafts, like limbs, are especially sensitive to this damage and can survive only 4-6 hours using traditional ice static cold storage (SCS). However, researchers recently demonstrated the superiority of an active form of storage outside of the body, ex vivo, and by perfusing a limb it could survive normally irreversible  damage from storage up to 12 hours (Kueckelhaus, et al.).

Professor Bohdan Pomahac’s lab at Brigham and Women’s Hospital recently published a paper evaluating changes in gene expression associated with this damage—particularly expression of genes involved in the response to lack of oxygen and nutrients (Krezdor, et al.). The researchers studied these genes by measuring RNA amounts from time point muscle biopsies of dissected Yorkshire female pig limbs placed in either SCS or ex vivo perfusion systems. They found that destructive pathways activated in response to low levels of oxygen and nutrients were significantly lower in perfused limbs vs traditional SCS limbs.

Altogether, these data suggest that the underlying benefits of ex vivo perfusion are conferred by slowing the expression of harmful genes, findings that may translate to other types of transplantable tissue. - Mitch Rostad

 

Freeze-drying: not just for astronauts’ ice cream

Clinical applications of tissue engineered products hold much promise, as grafts derived from or repopulated with a patient’s own cells (or cells modified to appear to be the patient’s) can alleviate many of the complications from rejection or immunogenicity of a graft. However, these products need be made available “off-the-shelf” if they are to be widely adopted and thus require preservation.

In a recent paper from the Gerosa lab out of Padua University Medical School, Zouhair et al. compared three methods of preservation head to head: slow-freezing, vitrification, and freeze-drying to preserve decellularized bovine pericardial (DBP) scaffolds (utilized in cardiac surgery to repair damaged heart valves or as a vascular grafts). While scaffolds that were slow-frozen and then rewarmed were weak and brittle when examined, both the vitrified and freeze-dried scaffolds had similar extracellular matrix structure, biomechanical properties and cellular reconstitution when compared to each other and a fresh scaffold. The authors conclude that more exploration should be done to tissue-banking via dry-preservation, as it can be more cost-effective than vitrification. - Dr. Kate Franz

 
 
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Jump to: OPA updates or Selected publications

Help keep the community updated by sending your news to alyssa.ward@organpreservationalliance.org

 

Recruiting two PhD students for preservation research project (Hannover, Germany)

Prof. Dr. med. vet. Harald Sieme and Prof. Dr. Ir. Willem F. Wolkers are recruiting two PhD students to work on a German Research Foundation-funded project to develop novel preservation strategies in reproductive and regenerative medicine. Click here for more info and application instructions!

 

OPA advisors haven’t been hibernating this winter…

  • Drs. Jason Acker and Janet Elliot jointly elucidated the effects of substrate thermal expansion on the ability for a monolayer of cells to remain adherant throughout freezing.

  • As explored in more detail above, Dr. Janet Elliott further developed a protocol to cryopreserve endothelial cell monolayers. The Elliott lab also explored the dependence of fluid evaporation on container geometry.

  • Dr. Greg Fahy and collaborators identified some of the genes behind cryoprotectant toxicity by mutagenizing cells and selecting for cryoprotective agent tolerance.

  • Dr. John Bischof characterized a laser gold nanowarming approach and collaborated to apply vitrification and laser warming to the preservation of coral larvae for conservation and research efforts.

  • OPA Advisors Drs. Mehmet Toner and Korkut Uygun collaborated to develop a novel approach to vitrify large droplet volumes.

  • Drs. Korkut Uygun and James Markmann showed that increased oxygen in preservation solution sustains ATP levels longer that traditional preservation solution, resulting in less ischemic damage.

  • Drs. James Markmann and David Sachs doubled the survival time of baboons in a model of post-hepatectomy liver failure using a swine xenograft,

  • Dr. David Sachs determined the lipid profiles of the longest surviving recipients of pig-to-primate liver xenotransplants.

  • Dr. Kenneth Storey purified a freeze-responsive enzyme from Rana sylvatica, studied the temperature-dependent post-translational modification of a metabolic enzyme, and wrote about how mammalian warm hibernators can provide valuable lessons for extending preservation times of human organs.

  • Dr. Bohdan Pomahac showed that perfusion of limbs reduces activation of hypoxia-related genes in a porcine model (click here to jump to the longer paper highlight).

  • Dr. Dayong Gao developed a technique for high-density, three-dimensional hepatocyte culture.

  • Dr. Boris Rubinsky explored the potential to preserve Escherichia coli isochorically.

 

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