Hypothermic Storage and Cryopreservation of Corneas

描述(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)的新技术。HypoThermosol(HTS)平台包括HTS-PURGE、HTS-BASE、HTS-DCC、HTS-FRS以及再生医学市场客户正在使用的其他定制保存解决方案。BioLife的无血清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，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，这些事件可能由于保存储存而启动。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的几个HTS变种已经被证明比目前设计用于器官存储的商业解决方案(例如ViaSpan)有明显的好处。BioLife的CryoStor系列在冷冻保存方面可能比目前的ATCC工业协议更好。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。BioLife已经认识到，由Chron Vision分销的角膜存储解决方案OptisoI-GS是在12年前开发的，其配方不是基于保存过程中角膜细胞发生的分子事件。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，这些技术将提供比OptisoI-GS更好的性能。第一阶段项目建议：(1)建立细胞培养方法和多终点分析系统，以评估体外培养的牛和人角膜细胞及其对HTS和CryoStor变异体的反应；(2)确定细胞凋亡和坏死在保存诱导的角膜细胞死亡中的相对贡献；(3)确定添加硫酸软骨素是否对HTS-Coral和CryoStor-Coral在低温下维持细胞和组织保护是必要的；以及(4)测试作为HTS-Coral和CryoStor-Corna添加剂的基质软骨素是否必要。第二阶段项目将测试HTS-角膜和冷冻-角膜对完整人类角膜的保存情况。

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.