Tracking and ablating subpopulations of epiblast cells in the chick embryo.

Tracking and ablating subpopulations of epiblast cells in the chick embryo.
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DOI:
10.1251/bpo145
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发表时间:
2008-09-01
影响因子:
6.4
通讯作者:
George-Weinstein M
George-Weinstein M
中科院分区:
生物学3区
文献类型:
--
作者:
Gerhart J;Neely C;Pfautz J;George-Weinstein M

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早期鸡胚胎包含表达谱系特异性转录因子的细胞亚群。我们已经制定了检查这些细胞在发育过程中的作用的方案,包括标记它们以用于细胞跟踪目的,并在上胚层内消融它们。该程序利用了这样一个事实,即上皮细胞亚群不同地表达由单抗识别的细胞表面抗原。胚胎被从蛋壳中取出,并与抗体一起孵化在蛋黄上。结合抗体的细胞要么用荧光二抗标记,要么用补体裂解。对于长期分析,胚胎被送回宿主壳并放入孵化器中。这种全胚胎操作、体外培养和体外孵化的方法支持正常发育到胎儿期。
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.
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期刊: POULTRY SCIENCE
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