ES CELL DERIVED LINEAGES--A NEW ENABLING TECHNOLOGY

描述(改编自申请人的摘要)：胚胎干细胞提供了一种新的 以及生物医学研究所需的资源。申请者建议 生成使能技术，帮助实现这一目标的全部潜力 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、淋巴样细胞、 心肌、骨骼肌和神经细胞。为了充分利用该系统，它 从胚胎干细胞开始，在体外获得完全纯净的血统是必不可少的 细胞。申请者将生成一组特定于以下各项的单抗 早期胚胎的主要谱系和识别细胞表面抗原。 利用这些单抗，申请者将对培养的ES细胞进行分选，以便 建立外胚层、中胚层或内胚层细胞的纯化培养。在一个 同时，申请者将提纯中胚层前体 利用胚胎干细胞遗传学。基因打靶将被用来产生一个 新霉素抗性基因插入矮秆基因的ES系 在早期中胚层谱系中特异表达。利用这条线路， 申请者将产生早期中胚层细胞的培养。使能 此应用程序产生的工具提供了迈向 未来的资源由大量遗传基因的储存库组成 基因工程的ES细胞系。

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.