DEVELOPMENT OF CRYOPLATES FOR CYTOTOXICITY TESTING

用于细胞毒性测试的低温板的开发

• 批准号: 6074687
• 负责人: LIA H CAMPBELL
• 金额: $ 10.03万
• 依托单位: ORGAN RECOVERY SYSTEMS, INC.
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-09-30 至 2001-09-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6074687
• 关键词: corneal endothelium; cryopreservation; cytotoxicity; evaluation /testing; extracellular matrix; technology /technique development; tissue /cell culture

The search for alternative methodologies to reduce the number of animals used in biomedical research and toxicology testing has focused on in vitro methods of cultured cells. Our concept is to develop an off-the-shelf frozen microtiter plate ("Cryoplate") which will provide a readily available cell delivery system for in vitro assaying of cytotoxicity. We propose to examine the general hypothesis that cells dispensed in a tissue culture plate and attached to either a natural or synthetic substrate can withstand the rigors of freezing and thawing to provide a cell delivery system for in vitro cytotoxicity assays. Initial studies will utilize cultured bovine corneal cells which permit convenient examination of tolerance to the stresses of cryopreservation, either as individual cells attached directly to the synthetic substrate of the tissue culture plate or as a layer of cells attached to the natural extracellular matrix (ECM) laid down by these cells in culture. It is envisaged that optimization of cryobiological variables will enable development of the Cryoplate concept for use in a wide variety of toxicology and diagnostic laboratories. This product will not only eliminate the need for plating and expanding cells for in vitro testing purposes, but will possibly require handling fewer cells than conventional methods. The basic technology developed in this research may eventually lead to the cryopreservation of cells on other substrates such as miniaturized synthetic chips carrying sensors or other diagnostic tools. PROPOSED COMMERCIAL APPLICATION: It is anticipated that cryopreserved cell delivery systems will be commercialized for chemical testing and diagnostic assays. The methods may be applicable to the development of in vitro cell assay kits for a variety of cell types and purposes. Licensing opportunities will be sought with industry partners for alternative applications.

寻找替代方法以减少生物医学研究和毒理学测试中使用的动物数量的研究重点集中在体外培养细胞方法上。我们的理念是开发一种现成的冷冻微量滴定板（“Cryoplate”），它将为体外细胞毒性测定提供易于使用的细胞输送系统。 我们建议检查一般假设，即分配在组织培养板中并附着在天然或合成基质上的细胞可以承受严酷的冷冻和解冻，从而为体外细胞毒性测定提供细胞递送系统。 初步研究将利用培养的牛角膜细胞，可以方便地检查对冷冻保存压力的耐受性，无论是直接附着在组织培养板的合成基质上的单个细胞，还是附着在天然细胞外基质（ECM）上的一层细胞由这些细胞在培养物中形成。 预计冷冻生物学变量的优化将使冷冻板概念的发展能够用于各种毒理学和诊断实验室。该产品不仅无需为体外测试目的而进行铺板和扩增细胞，而且可能比传统方法需要处理更少的细胞。 这项研究开发的基础技术最终可能会导致细胞在其他基质上的低温保存，例如携带传感器或其他诊断工具的小型合成芯片。拟议的商业应用：预计冷冻细胞输送系统将商业化用于化学测试和诊断测定。该方法可适用于开发用于多种细胞类型和目的的体外细胞测定试剂盒。 将与行业合作伙伴寻求替代应用的许可机会。