Hypothermic and Cryopreservation of Human Blood Vessels

描述(申请人提供)：BioLife Solutions Inc是一家生物包装公司，致力于为人类细胞和组织的低温存储(4至8摄氏度)和低温保存(-196摄氏度)开发改进的解决方案。HTS平台包括HTS-BASE、HTS-DCC、HTS-FRS以及目前正在为再生医学市场的客户设计的其他定制保存解决方案。BioLife的CryoStor冷冻保存解决方案平台专为在液氮中存储细胞和组织而设计，无需血清，并降低了DMSO水平。BioLife的解决方案是通过首先了解细胞死亡级联的分子细节来设计的，这些级联可能会因低温储存或低温保存而过早启动。一旦知道，溶液配方就会改变，从而抑制这些细胞死亡过程。BioLife的HTS-FRS优于UW溶液(Viaspan)-目前用于大多数人体器官存储的溶液。同样，BioLife的CryoStor系列在冷冻保存方面可能比目前的ATCC行业协议更好。BioLife的解决方案现已成为生物心脏FDA批准的细胞心肌成形术IND的一部分，这是心脏再生医学领域的一项成功的国际计划，部分原因是BioLife的HTS-FRS。这项第二阶段的提案致力于改进人类血管的低温和冷冻保存。BioLife公司与保存的同种异体血管假体的主要供应商CryoLife公司合作，从而可以开发改进的保存解决方案，以(1)不需要目前包括血清和硫酸软骨素，以及(2)利用BioLife公司最近关于细胞凋亡在保存失败中的作用的研究，从而改善保存的同种移植血管的移植结果。这项建议的具体目标包括：(1)确定储存在HTS和CryoStor溶液中的细胞存活能力的提高是否反映出与Viaspan类似的功能改善；(2)测试是否包括其他调节细胞凋亡的药物，看它们是否具有改善HTS和CryoStor疗效的潜在能力；(3)确定应用Ciphergen蛋白质芯片系统是否可用于高通量评估HTS和CryoStor变异体和/或确定保存的血管是否受到细菌肠毒素的污染；以及(4)测试可能使细胞具有异常低温韧性的自然因素。

DESCRIPTION (provided by applicant): 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.