KIT FOR CONSTRUCTING ES CELL/BLASTOCYST CHIMERAS

用于构建 ES 细胞/囊胚嵌合体的试剂盒

• 批准号: 2728553
• 负责人: JOHN D BIGGERS
• 金额: $ 37.91万
• 依托单位: CELL AND MOLECULAR TECHNOLOGIES, INC.
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-09-01 至 2000-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2728553
• 关键词: biomedical equipment development; early embryonic stage; embryo /fetus culture; embryo /fetus preservation; embryo /fetus transplantation; embryonic stem cell; growth media; laboratory mouse; tissue mosaicism

DESCRIPTION: The aim of this proposal is to produce a cost-effective kit that will facilitate the production of mouse embryonic stem (ES) cell / blastocyst chimeras to make genetically engineered mice. The kit will be designed so that it can be used by investigators with only limited experience with mouse embryo culture and embryo transfer. Costs of making knock-out mice will be significantly reduced by eliminating the need for local animal colonies for the production of 8-cell mouse embryos by supplying them from a central stock of frozen embryos. The thawed 8-cell embryos will be aggregated with the ES cells and then transferred to the uteri of pseudopregnant surrogate mothers. In Phase II of this application, the following research will be done to produce an optimized kit: (1) a method will be developed to assay rapidly the aggregation of ES cells with 8-cell embryos; (2) KSOM/AA, a new medium that is very effective for the culture of mouse preimplantation embryos, will be optimized for the aggregation of ES cells and mouse embryos; (3) the culture of ES cells on a feeder layer will be improved by replacing the feeder layer with a plate of supporting killed primary mouse embryonic fibroblasts; (4) KSOM/AA will be modified to replace Dulbecco's MEM for the culture of ES cells; and (5) data will be collected to determine which of two inbred strains of mouse (C57BL/6 or FVB/N) are preferable as donors of the 8-cell embryos used for aggregation. PROPOSED COMMERCIAL APPLICATION: NOT AVAILABLE

描述：该提案的目的是生产一个具有成本效益的套件 这将促进小鼠胚胎茎（ES）细胞的产生 胚泡嵌合体制成基因工程的小鼠。 套件将是 设计以便仅有限的调查员可以使用它 具有小鼠胚胎培养和胚胎转移的经验。 制作成本 消除对淘汰小鼠的需求将大大减少 当地动物菌落生产8细胞小鼠胚胎 从冷冻胚胎的中央库存中供应它们。 解冻的8细胞 胚胎将与ES细胞聚集，然后转移到 伪孕妇代理母亲的子宫。 在第二阶段 应用，将进行以下研究以产生优化 套件：（1）将开发一种方法来快速测定聚集 ES具有8细胞胚胎的细胞； （2）KSOM/AA，一种非常非常的新媒介 对小鼠植入前胚胎的培养有效，将是 针对ES细胞和小鼠胚胎的聚集进行了优化； （3） 通过替换将改善ES细胞在馈线上的培养 带有一盘支撑杀死的主小鼠胚胎的馈线层 成纤维细胞； （4）KSOM/AA将被修改以替换Dulbecco的MEM ES细胞的培养； （5）将收集数据以确定 两种近交小鼠菌株（C57BL/6或FVB/N）是捐助者的优选 用于聚集的8细胞胚胎。 拟议的商业应用程序：不可用