Hypothermic Storage and Cryopreservation of Corneas

描述（由申请人提供）BioLife Solutions Inc是一家专门从事生物包装的公司，致力于开发人体细胞和组织低温储存（4至8 ℃）和冷冻保存（-196 ℃）的新技术。HypoThermosol（HTS）平台由HTS-PURGE、HTS-BASE、HTS-DCC、HTS-FRS和其他定制保存解决方案组成，供再生医学市场的客户使用。BioLife的无血清CryoGlycoprotein冷冻保存溶液平台专为液氮中的细胞和组织储存而设计，需要降低DMSO水平。BioLife的解决方案是围绕对细胞死亡级联反应的分子事件的理解而设计的，这些细胞死亡级联反应可以由于保存储存而启动。一旦已知，溶液配方以抑制这些细胞死亡过程的方式改变。BioLife的几种HTS变体已被证明比目前设计用于器官储存的商业解决方案（例如ViaSpan）具有显着的益处。BioLife的冷冻系列可能比目前的ATCC行业协议更好地进行冷冻保存。BioLife的解决方案现在是BioHeart的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是一种由Chiron Vision分销的角膜储存溶液，是12年前开发的，其配方不是基于保存角膜细胞发生的分子事件。根据SBIR计划，BioLife将开发专门的角膜保存技术，HTS-CORNEA和CryoStor-CORNEA，这将提供优于OptisoI-GS的性能。第一阶段项目提出：（1）开发一种细胞培养方法和多终点测定系统，以评估体外牛和人角膜细胞及其对HTS和冷冻变体的反应;（2）确定细胞凋亡和坏死在冷冻诱导的角膜细胞死亡中的相对贡献;（3）确定加入硫酸软骨素（存在于OptisoI-GS和其他角膜储存溶液中的细胞外基质组分分子），是HTS-CORNEA和CryoStor-CORNEA在寒冷中维持细胞和组织保护所必需的;和（4）测试MMP和NO抑制剂作为添加剂包含到HTS-CORNEA和CryoStor-CORNEA中。第二阶段项目将测试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.