SBIR-STTR Award

BioLife Technologies (BLT) was incorporated on March 31, 1998. BLT's primary mission is to develop improved hypothermic and cryopreservation solutions designed to maintain cells, tissues and organs in a near state of suspended animation. Cardioplegia, organ transplant and selected non- regulated markets have been targeted for development. This Phase I SBIR proposes to use four cardiovascular cell strains as in vitro tools to help develop a future HTS solution that is optimized for human blood vessel preservation. BLT's hypothermic solutions, the HypoThermosol(R) (HTS) series, are better at cold-protecting kidney, heart and skin cells than is ViaSpan(R) - a product produced by DuPont-Merck ten years ago. ViaSpan(R) currently commands the largest market share in the preservation solution business, but the design of ViaSpan(R) is not based on modern molecular biology investigations. BLT has launched an aggressive program to determine the molecular basis of cell death during extended hypothermic and cryopreservation storage so that knowledge of these events can lead to a new generation of hypothermic solutions. As a result of BLT's early success, HTS is currently being used in a pancreatic islet transplantation Phase I clinical trial. DNA gels presented in this proposal demonstrate that cells cold stored for too long or cryopreserved die by apoptosis (programmed cell death) two days after they are returned to normothermic (37 degrees C) temperatures. HTS supplemented with apoptosis inhibitors blocks this process and improves HTS's performance. BLT currently has a patent pending that will allow BLT the exclusive right to formulate future HTS solutions by design so that apoptosis is inhibited. The Specific Aims of this Phase I project are to (a) determine the relative contributions of apoptosis and necrosis as a consequence of preservation-induced cellular injury to blood vessel cells; (b) determine which apoptosis inhibitors are the most effective at increasing HTS efficacy; (c) determine the relationship, if any, between necrosis/apoptosis, cytochrome C leakage from mitochondria, and loss of mitochondrial transmembrane potential; and (d) determine if extended preservation shortens telomeres and as a consequence precipitates premature cell senescence. Phase II studies will be devoted to HTS preservation of intact human blood vessels - work to be accomplished with our collaborator, CryoLife Inc. PROPOSED COMMERCIAL APPLICATIONS: BioLife is developing a new generation of preservation solutions that will improve the long term storage of human blood vessels used for clinical cardiovascular applications. The solutions will also facilitate preservation of future, tissue engineered blood vessels currently under development.

Thesaurus Terms:
blood vessel, cryopreservation, cryoprotective agent, genetic strain, programmed cell death, technology /technique development, tissue /organ preservation coronary artery, freezing, hypothermia, membrane potential, necrosis, telomerase, vascular smooth muscle

BioLife Solutions Inc is a Biological Packaging company dedicated to developing improved solutions for hypothermic storage (4 to 8C) and cryopreservation (-196C) of human cells and tissues. The HTS platform consists of HTS-BASE, HTS-DCC, HTS-FRS and other custom preservation solutions currently being designed for clients in the Regenerative Medicine market. BioLife's CryoStor cryopreservation solution platform is designed for storage of cells and tissues in liquid nitrogen without serum and with reduced levels of DMSO. BioLife's solutions are designed by first understanding the molecular details of cell death cascades that can be prematurely launched as a consequence of hypothermic storage or cryopreservation. Once known, solution formulation is altered in such a manner that inhibits these cell death processes. BioLife's HTS-FRS is superior to UW Solution (ViaSpan) - the solution currently being used for most human organ storage. Similarly, BioLife's CryoStor series may be better at cryopreservation than the current ATCC industry protocol. BioLife's solutions are now a part of BioHeart's FDA-approved IND in cellular cardiomyoplasty - a successful international program in cardiac Regenerative Medicine made possible, in part, by BioLife's HTS-FRS. This Phase II proposal is dedicated to the improved hypothermic and cryopreservation of human blood vessels. BioLife has teamed with CryoLife Inc., a major supplier of preserved, allograft vascular prostheses, so that improved preservation solutions can be developed that (1) do not necessitate the current inclusion of serum and chondroitin sulfate and (2) harness BioLife's recent research on the role of apoptosis in preservation failure so that outcome is improved in the transplant of preserved allograft vessels. The Specific Aims of this proposal include the following: (1) Determine if the improvement in viability of cells stored in HTS and CryoStor solutions relative to ViaSpan is mirrored by a similar improvement in function; (2) Test the inclusion of other agents that modulate apoptosis for their potential abilities to improve HTS and CryoStor efficacy; (3) Determine if applying the Ciphergen ProteinChip system can be used for either high throughput assessment of HTS and CryoStor variants and/or determining whether preserved vessels are contaminated with bacterial enterotoxins and (4) test for natural factors that may confer unusual hypothermic resilience in cells.

Thesaurus Terms:
apoptosis, blood vessel, cryopreservation, cryoprotective agent, hypothermia, technology /technique development, tissue /organ preservation chondroitin sulfate, cryoscience, serum, vascular smooth muscle human tissue, mass spectrometry, microarray technology