HTS/CryoStor-Liver- Is Viability Linked to Function?

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平台由HTS-BASE、HTS-DCC、HTS-FRS和定制低温保存解决方案组成，目前正在为再生医学市场的客户设计。BioLife的低温保存解决方案平台是为在液氮中存储细胞和组织而设计的。BioLife的解决方案是通过了解细胞死亡级联的分子细节来设计的，这些细节可以因低温储存或低温保存而被激活。一旦了解，就会改变溶液配方，从而抑制这些细胞死亡过程。作为这种合理的分子方法的结果，BioLife的HTSFRS优于目前用于低温保存大多数人体器官以供移植的LIW溶液(Viaspan)。同样，BioLife的无血清CryoStor系列可能比目前的ATCC行业协议更适合用于冷冻保存。BioLife的解决方案现已获得美国食品和药物管理局批准，作为BioHea_‘S公司细胞心肌成形术的一部分。这一成功引起了组织转化技术公司(T3)的兴趣，该公司计划利用原代人类肝细胞进行细胞治疗应用，以治疗肝脏疾病。BioLife开发了一种HTS-Liver原型平台，它可以保持人类肝癌细胞系的活力，是UW的几倍。CryoStor数据库的开发表明，也有可能开发一个CryoStor-Liver平台。这项建议的具体目的是确定：(1)HTS-肝对原代人类肝细胞的作用是否与其对人肝癌细胞系的作用相同；(2)低温保存的原代人肝细胞的存活率和功能是否存在联系；(3)对HTS-肝脏的碱基进行修改后，同样可以导致低温保存肝的发育；(4)对HTS-肝或低温保存肝的修饰可以提高它们在保存过程中保护细胞生存和功能的能力。高温超导肝和低温冷冻肝的研制是支持T3‘S人肝细胞治疗在再生医学中应用的关键。此外，改进的低温和低温保存平台的开发将立即使T3受益，因为它提供了一种手段，使原代人类肝细胞进入更大的国内和国际研究市场。

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 custom hypothermic 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. BioLife's solutions are designed by f_ understanding the molecular details of cell death cascades that can be activated as a consequence of hypothermic storage or cryopreservation. Once understood, solution formulation is altered such that these cell death processes are inhibited. As a consequence of this rational, molecular approach, BioLife's HTSFRS is superior to LIW Solution (ViaSpan), the solution currently used for hypothermic preservation of most human organs for transplantation. Similarly, BioLife's serum-free CryoStor series may be better for cryopreservation than the current ATCC industry protocol. BioLife's solutions are now FDA-approved as part of BioHea_'s IND in cellular cardiomyoplasty. This success captured the interest of Tissue Transformation Technologies (T3), which plans cell therapy applications with primary human hepatocytes to treat liver disease. BioLife developed a prototype HTS-LIVER platform that preserves the viability of a human hepatoma cell line several-fold longer than UW. The development of CryoStor-BASE suggests that the development of a CryoStor-LIVER platform is also probable. The Specific Aims of this proposal are to determine if: (1) HTS-LIVER works equally as well with primary human hepatocytes as it does with a human hepatoma cell line; (2) viability and function of hypothermically preserved primary human hepatocytes are linked; (3) the same modifications to HTS-BASE that generated HTS-LIVER can result in the development of CryoStor-LIVER; (4) modifications to HTS-LIVER or CryoStor-LIVER can improve their abilities to protect cell viability and function during preservation. Development of HTS-LIVER and CryoStor-LIVER are critical to support T3's human hepatocyte cell therapy applications in Regenerative Medicine. In addition, development of improved hypothermic and cryogenic preservation platforms will immediately benefit T3 by providing a means to reach larger national and international research markets with primary human hepatocytes.