Hypothermic Storage and Cryopreservation of Corneas

BioLife Solutions Inc .是一家专业从事生物包装的公司，致力于开发人体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）的新技术。低温溶胶（HTS）平台包括HTS- purge、HTS- base、HTS- dcc、HTS- frs和其他定制保存解决方案，正在再生医学市场的客户中使用。BioLife的无血清CryoStor冷冻保存溶液平台专为细胞和组织在液氮中储存而设计，需要降低DMSO水平。BioLife的解决方案是围绕着对细胞死亡级联的分子事件的理解而设计的，这些事件可能是由于保存存储而启动的。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的几种HTS变体已被证明比目前为器官储存而设计的商业解决方案（例如viaspan）具有明显的优势。BioLife的CryoStor系列在低温保存方面可能比目前的ATCC工业协议更好。BioLife的解决方案现在是BioHeart获得fda批准的细胞心肌成形术IND的一部分，这是一个成功的心脏再生医学国际项目，部分原因是BioLife的HTS-FRS。BioLife已经认识到，OptisoI-GS，一种由Chiron Vision销售的角膜储存液，是12年前开发的，其配方不是基于角膜细胞在保存过程中发生的分子事件。根据这项SBIR计划，BioLife将开发专门的角膜保存技术，HTS-CORNEA和CryoStor-CORNEA，这将提供比OptisoI-GS更好的性能。I期项目建议：(1)开发一种细胞培养方法和多终点检测系统，以评估体外培养的牛和人角膜细胞及其对HTS和CryoStor变体的反应；(2)确定保存诱导的角膜细胞死亡中凋亡和坏死的相对贡献；(3)确定是否需要添加硫酸软骨素（OptisoI-GS和其他角膜储存液中存在的细胞外基质成分分子）来维持HTS-CORNEA和CryoStor-CORNEA在低温下对细胞和组织的保护；(4)检测MMP和NO抑制剂作为HTS-CORNEA和CryoStor-CORNEA添加剂的掺入情况。二期项目将测试HTS-Cornea和CryoStor-Cornea对整个人类角膜的保存。

DESCRIPTION (provided by applicant) BioLife Solutions Inc is a company specializing in Biological Packaging and dedicated to developing new technologies for hypothermic storage (4 to 8C) and cryopreservation (-196C) of human cells and tissues. The HypoThermosol (HTS) platform consists of HTS-PURGE, HTS-BASE, HTS-DCC, HTS-FRS and other custom preservation solutions being utilized by clients in the Regenerative Medicine market. BioLife's serum-free CryoStor cryopreservation solution platform is designed for cell and tissue storage in liquid nitrogen and requires reduced DMSO levels. BioLife's solutions are designed around an understanding of the molecular events underlying cell death cascades that can be launched due to preservation storage. Once known, solution formulation is altered in such a manner that inhibits these cell death processes. Several of BioLife's HTS variants have been shown to have a marked benefit over present commercial solutions (e.g.ViaSpan) designed for organ storage. 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. BioLife has recognized that OptisoI-GS, a corneal storage solution distributed by Chiron Vision, was developed 12 years ago and its formulation is not based on the molecular events that occur to corneal cells subjected to preservation. Under this SBIR program, BioLife will develop specialized corneal preservation technologies, HTS-CORNEA and CryoStor-CORNEA, that will provide enhanced performance over that of OptisoI-GS. The Phase I project proposes to: (1) Develop a cell culture method and multiple endpoint assay system to assess bovine and human corneal cells in vitro and their responses to HTS and CryoStor variants; (2) Determine the relative contributions of apoptosis and necrosis in preservation-induced, corneal cell death; (3) Determine if the addition of chondroitin sulfate, an extracellular matrix component molecule present in OptisoI-GS and other corneal storage solutions, is necessary for HTS-CORNEA and CryoStor-CORNEA to maintain cell and tissue protection in the cold; and (4) Test the inclusion of MMP and NO inhibitors as additives to HTS-CORNEA and CryoStor-CORNEA. The Phase II project will test HTS-Cornea and CryoStor-Cornea on the preservation of whole, human corneas.