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

描述（由申请人提供）：BioLife Solutions Inc .是一家生物包装公司，致力于开发人类细胞和组织的低温储存（4至8摄氏度）和低温保存（-196摄氏度）的改进解决方案。HTS平台包括HTS- base、HTS- dcc、HTS- frs以及目前正在为再生医学市场客户设计的定制低温保存解决方案。BioLife的CryoStor冷冻保存溶液平台是专为在液氮中储存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而设计的，这种级联可能由于低温储存或低温保存而被激活。一旦理解，溶液配方被改变，这些细胞死亡过程被抑制。由于这种合理的分子方法，BioLife的HTSFRS优于LIW溶液（ViaSpan）， LIW溶液目前用于大多数移植用人体器官的低温保存。同样，BioLife的无血清CryoStor系列可能比目前的ATCC工业协议更适合冷冻保存。BioLife的解决方案现已获得fda批准，作为BioHea_细胞心肌成形术IND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肝细胞进行细胞治疗，以治疗肝脏疾病。BioLife开发了一种原型HTS-LIVER平台，该平台保留了人类肝癌细胞系的生存能力，比UW长几倍。CryoStor-BASE的发展表明，CryoStor-LIVER平台的发展也是可能的。本提案的具体目的是确定：(1)HTS-LIVER对原代人肝细胞的作用是否与对人肝癌细胞系的作用一样好；(2)低温保存的原代人肝细胞的活力和功能是相互关联的；(3)生成HTS-LIVER的HTS-BASE的相同修改可以导致CryoStor-LIVER的开发；(4)对HTS-LIVER或CryoStor-LIVER进行修饰可以提高其在保存过程中保护细胞活力和功能的能力。HTS-LIVER和CryoStor-LIVER的发展对于支持T3在再生医学中的人肝细胞治疗应用至关重要。此外，改进的低温和低温保存平台的发展将立即使T3受益，为原代人肝细胞进入更大的国内和国际研究市场提供了一种手段。

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.