KIT FOR CONTRUCTING ES CELL/BLASTOCYST CHIMERAS

用于构建 ES 细胞/囊胚嵌合体的试剂盒

• 批准号: 2418085
• 负责人: JOHN D BIGGERS
• 金额: $ 10万
• 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 1998-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2418085
• 关键词: biomedical equipment development; embryo /fetus culture; embryo /fetus preservation; embryo /fetus transplantation; growth media; laboratory mouse

DESCRIPTION: (Adapted from the Applicant's Abstract): The aim of this proposal is to produce a cost effective kit that will facilitate the production of mouse embryonic stem (ES) cell/chimeras to produce genetically engineered mice. The kit will be designed for use by investigators with only limited experience with mouse embryo culture and embryo transfer. Costs will be 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 into the uteri of pseudopregnant surrogate mothers. Improvement of the embryo culture systems used in the several steps of the procedure will be made by optimizing the culture medium KSOM AA/Gluc, a new very effective culture medium for preimplantation mouse embryos. The feasibility study in this Phase I application will test the following hypotheses: (1) KSOM AA/Gluc can replace Dulbecco's modification of Eagle's minimum essential medium for the culture of embryonic stem cells, (2) Thawed, previously frozen, 8-cell embryos, cultured in KSOM AA/Gluc develop into blastocysts that can then develop into normal fetuses after transfer to the uterus of surrogate mothers, and (3) Aggregates of embryonic stem cells with thawed, previously frozen, 8-cell embryos, cultured in KSOM AA/Gluc, can develop into newborn mice that are embryonic stem cell/blastocyst derived chimeras.

描述：（根据申请人的摘要进行了改编）： 建议是生产一个具有成本效益的套件，以促进 产生小鼠胚胎茎（ES）细胞/嵌合体以产生 基因工程小鼠。 该套件将设计供 对小鼠胚胎培养和有限经验的研究人员 胚胎转移。 消除对本地的需求，将降低成本 通过供应生产8细胞小鼠胚胎的动物菌落 它们来自冷冻胚胎的中央库存。 解冻的8细胞胚胎 将与ES细胞聚合，然后转移到子宫中 伪孕妇代理母亲。 改善胚胎文化 该过程的几个步骤中使用的系统将由 优化培养基KSOM AA/GLUC，一种新的非常有效的文化 植入前小鼠胚胎的培养基。 此中的可行性研究 第一阶段应用将测试以下假设：（1）KSOM AA/GLUC 可以取代Dulbecco修改Eagle的最低必需媒介 对于胚胎干细胞的培养，（2）融化，以前冷冻， 8细胞胚胎，在ksom aa/gluc中培养的胚胎发展成胚泡， 然后可以在转移到子宫后发展成正常胎儿 替代母亲和（3）胚胎干细胞的聚集体 在ksom aa/gluc中培养的融化，以前冷冻的8细胞胚胎可以 发展成胚胎干细胞/胚泡的新生小鼠 嵌合体。