December 2018
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!
Jump to: Selected publications or News from our network
This June, the American Society of Transplantation Recovery and Preservation Community of Practice (RAP COP) hosted a workshop, in which participants requested further efforts dedicated to advancing machine perfusion. OPA and RAP COP are, therefore, proud to announce the launch of a new working group doing just that.
This group, including top executives from Organ Procurement Organizations and machine perfusion companies, leading researchers and clinicians, and dedicated advocates, represents a powerful collaboration between stakeholders dedicated to leveraging advances in machine perfusion technology to save and improve lives.
We held a kick-off meeting to set our 2019 priorities and it was fantastic to see so much expertise generating such promising ideas. Ultimately we plan to focus on (1) FDA outreach and education; (2) Standardizing nomenclature and defining important metrics with the goal of using the established common vocabulary to furnish a database of information about and outcomes of transplants with perfused organs; (3) Implementation logistics and (4) Developing a network of scientists that use perfusion for biomedical research - whether working on machine perfusion or using it as a platform for gene modification, drug delivery, organ preservation, etc.
If you’d like to get involved, email alyssa.ward@organpreservationalliance.org!
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.
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.
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.
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!
Many cell types are regularly frozen in suspension with reasonably high cell viabilities. However, without the ability to preserve cell-cell interactions and higher order structures, these techniques fail to translate to engineered tissues, xenografts, and human organs. Professor Janet Elliott's lab at the University of Alberta sought to address this by optimizing a protocol to cryopreserve cells in a monolayer (Eskandari, et al.). They hypothesized that the presence of junctions between cells would increase ice formation inside the cells and would necessitate addition of a cryoprotective agent (CPA) that can get inside and prevent that damage.
By using a modified CPA cocktail aimed at preventing intracellular damage and precipitating ice formation prior to cooling they were able to obtain over 90% cell viability while slow freezing cells attached to a coated surface. The substrate used in the optimized protocol experiences thermal expansion similarly to ice, building on concurrent findings, also from a collaboration with the Elliott group, that substrate expansion contributes to loss of adhesion during preservation (Rutt, et al.). This effect of thermal expansion could prove important to engineering tissues that can withstand preservation. - Dr. Alyssa Ward
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
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
Jump to: OPA updates or Selected publications
Help keep the community updated by sending your news to alyssa.ward@organpreservationalliance.org
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!
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.
Would you like to be involved with Biopreservation Briefings? Want your news included? E-mail alyssa.ward@organpreservationalliance.org with questions, ideas, and updates!
