ES CELL DERIVED LINEAGES--A NEW ENABLING TECHNOLOGY

ES 细胞衍生谱系——一种新的使能技术

• 批准号: 2772049
• 负责人: DAVID I GOTTLIEB
• 金额: $ 15.49万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-09-05 至 2000-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2772049
• 关键词: biotechnology; cell differentiation; cell fusion; cell line; drug resistance; ectoderm; embryonic stem cell; endoderm; flow cytometry; gene targeting; genetically modified animals; hamsters; hybrid cells; hybridomas; laboratory mouse; mesoderm; monoclonal antibody; neomycin; surface antigens

DESCRIPTION (Adapted from the applicant's abstract): ES cells provide a new and needed resource for biomedical research. The applicants propose to generate enabling technology to help realize the full potential of this system. ES cells are capable of differentiating in vitro into a wide variety of cell types including hematopoietic, endothelial, lymphoid, cardiac, skeletal muscle, and neural cells. To fully exploit the system, it will be essential to obtain pure lineages entirely in vitro starting with ES cells. The applicants will generate a panel of mAbs that are specific to the major lineages of the early embryo and recognize cell surface antigens. Utilizing these mAbs, the applicants will sort cultured ES cells so as to establish purified cultures of ectoderm, mesoderm, or endoderm cells. In a parallel effort, the applicants will purify the mesodermal precursor utilizing ES cell genetics. Gene targeting will be employed to generate an ES line with a neomycin resistance gene inserted in the brachyury gene expressed specifically in early mesoderm lineages. Utilizing this line, the applicants will generate cultures of early mesoderm cells. The enabling tools resulting from this application provide an essential step towards a future resource consisting of a repository of large numbers of genetically engineered ES cell lines.

描述（改编自申请人的摘要）：ES细胞提供了一种新的 和生物医学研究所需的资源。 申请人建议 产生使能技术，以帮助实现这一 系统 ES细胞能够在体外分化为广泛的 多种细胞类型，包括造血细胞，内皮细胞，淋巴细胞， 心脏骨骼肌和神经细胞 为了充分利用该系统， 将是从ES开始在体外完全获得纯谱系所必需的 细胞 申请人将产生一组特异于 早期胚胎的主要谱系，并识别细胞表面抗原。 利用这些mAb，申请人将对培养的ES细胞进行分类，以便 建立外胚层、中胚层或内胚层细胞的纯化培养物。 中 平行的努力，申请人将纯化中胚层前体 利用胚胎干细胞遗传学。 将采用基因靶向来产生 在短尾畸形基因中插入新霉素抗性基因的ES系 在早期中胚层谱系中特异表达。 利用这条线， 申请人将产生早期中胚层细胞的培养物。 有利 从这一应用程序产生的工具提供了一个重要的一步， 未来的资源，包括大量的基因库， 工程化ES细胞系。