Cultured Human Cell Monolayers Cryopreserved In Situ

描述（由申请人提供）：细胞保存服务公司（CPSI）开发了细胞系统低温储存的新方法。CPSI的策略是围绕由于广泛的细胞应激而启动的细胞死亡级联的分子事件设计的。一旦知道，就会开发出调节这些细胞死亡过程和提高细胞存活率的方法。CPSI的科学家吗?最近参与开发一种新的冷冻保存介质（CryoStor），加上目前与保存介质公司（BioLife Solutions - CryoStor）和其他生物技术公司的联系，他们销售原代人肝脏和皮肤细胞，这促使CPSI对进一步探索原位冷冻保存方法和设备的兴趣，以大量冷冻（处理）含有人类细胞的多孔培养皿。拟议的调查建立在目前CPSI科学家自20世纪90年代末以来一直在进行的方法和研究的基础上。本提案的目的是实现以下目标：SA1 -确定用于原位冷冻保存的最佳载体培养基；SA2 -测试混合冷冻保护剂，以确定哪些能提供最大的冷冻保护；SA3 -测试不同的细胞粘附和培养基质在解冻后最大限度地粘附人细胞的能力；测试专有的细胞培养板，以确定适合该工艺的候选细胞。SA5-确定冷却速率，最大限度地提高细胞存活率；SA6吗?将原位保存的细胞活力和功能与使用传统悬浮技术保存的细胞进行比较。第二阶段将致力于扩大高通量筛选工艺（即384孔板，在-80℃下储存一年）。CPSI计划在第三阶段将开发的技术授权给小型生物技术公司和大型制药工业。

DESCRIPTION (provided by applicant): Cell Preservation Services Inc (CPSI) develops new approached for low temperature storage of cellular system. CPSI's strategy is designed around the molecular events underlying cell death cascades that are launched due to extensive cell stress. Once known, methods are developed which modulate these cell death processes and improve cell survival. CPSI's scientist?s recent involvement in developing a new cryopreservation media (CryoStor), coupled with current associations with preservation media companies (BioLife Solutions - CryoStor) and other biotech companies who distribute primary human liver and skin cells, has precipitated CPSI's interest in further exploring in situ cryopreservation methodologies and devices to bulk freeze (process) multiwell dishes containing human cells. The proposed investigations build upon approaches and studies that current CPSI scientists have been working on since the late-1990?s. The intent of this proposal is to accomplish the following: SA1 - Identify the best carrier medium to use as the cryopreservation solution for in situ cryopreservation; SA2 - Test a cocktail of cryoprotective agents to determine those that confer the maximum cryoprotection; SA3 - Test different cell adhesion and culture matrices for their abilities to maximize the adhesion of human cells subsequent to thawing; SA4 - Test proprietary cell culture plates to identify suitable candidates for the process. SA5- Identify cooling rates that maximize cell survival; SA6 ? Compare cell viability and function in cells preserved in situ to those preserved using conventional suspended technology. Phase II will be dedicated to scaling up the process for high throughput screening (i.e. 384 well plates, one year storage time at -80C). CPSI plans to license the developed technology to both small biotechnology companies and large pharmaceutical industries in Phase III.