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Tracking and ablating subpopulations of epiblast cells in the chick embryo

机译:跟踪和消融雏鸡胚中表皮细胞的亚群

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The early chick embryo contains subpopulations of cells that express lineage-specific transcription factors. We have developed protocols to examine the role of these cells during development that involve labeling them for cell tracking purposes and ablating them within the epiblast. The procedures take advantage of the fact that subpopulations of epiblast cells differentially express cell surface antigens recognized by monoclonal antibodies. Embryos are removed from the shell and incubated on the yolk with an antibody. Cells that bind the antibody are either tagged with a fluorescent secondary antibody or lysed with complement. For long-term analyses, embryos are returned to a host shell and placed in an incubator. This method of whole embryo manipulation ex-ovo and incubation in-ovo supports normal development into the fetal period.
机译:雏鸡早期胚胎包含表达谱系特异性转录因子的细胞亚群。我们已经开发出协议来检查这些细胞在发育过程中的作用,包括标记它们以进行细胞追踪并在表皮细胞中消融它们。该程序利用了以下事实:表皮细胞的亚群差异表达单克隆抗体识别的细胞表面抗原。从壳中取出胚胎,并在蛋黄中与抗体一起孵育。结合抗体的细胞用荧光二抗标记或用补体裂解。为了进行长期分析,将胚胎放回宿主壳中并置于培养箱中。这种从卵外进行全胚处理和卵内孵化的方法支持胎儿正常发育。

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