Etches R J, Clark M E, Toner A, Liu G, Gibbins A M
Department of Animal & Poultry Science, University of Guelph, Ontario, Canada.
Mol Reprod Dev. 1996 Nov;45(3):291-8. doi: 10.1002/(SICI)1098-2795(199611)45:3<291::AID-MRD5>3.0.CO;2-N.
Chicken blastodermal cells were cultured for 48 hr as explanted intact embryos, as dispersed cells in a monolayer, or with a confluent layer of mouse fibroblasts. The cells were then dispersed and injected into stage X (E-G&K) recipient embryos that were exposed to 600 rads of irradiation from a 60Co source. Regardless of the conditions in which the cells were cultured, chimeras with contributions to both somatic tissues and the germline were observed. When blastodermal cells were co-cultured with mouse embryonic fibroblasts, significantly more somatic chimeras were observed and the proportion of feather follicles derived from donor cells was increased relative to that observed following the injection of cells derived from explanted embryos or monolayer cultures. Culture of blastodermal cells in any of the systems, however, yielded fewer chimeras that exhibited reduced contributions to somatic tissues in comparison to the frequency and extent of somatic chimerism observed following injection of freshly prepared cells. Contributions to the germline were observed at an equal frequency regardless of the conditions of culture, but were significantly reduced in comparison to the frequency and rate of germline transmission following injection of cells obtained directly from stage X (E-G&K) embryos. These data demonstrate that some cells retain the ability to contribute to germline and somatic tissues after 48 hr in culture and that the ability to contribute to the somatic and germline lineages is not retained equally.
鸡胚盘细胞作为完整胚胎外植体培养48小时,或作为单层分散细胞培养,或与汇合的小鼠成纤维细胞层共同培养。然后将细胞分散并注射到经60Co源600拉德照射的X期(E - G&K)受体胚胎中。无论细胞培养的条件如何,均观察到对体细胞组织和生殖系均有贡献的嵌合体。当胚盘细胞与小鼠胚胎成纤维细胞共同培养时,观察到的体细胞嵌合体明显更多,并且与注射外植胚胎或单层培养物来源的细胞相比,来自供体细胞的毛囊比例增加。然而,与注射新鲜制备的细胞后观察到的体细胞嵌合频率和程度相比,在任何系统中培养胚盘细胞产生的对体细胞组织贡献减少的嵌合体更少。无论培养条件如何,对生殖系的贡献观察频率相同,但与直接从X期(E - G&K)胚胎获得的细胞注射后生殖系传递的频率和速率相比,显著降低。这些数据表明,一些细胞在培养48小时后仍保留对生殖系和体细胞组织的贡献能力,并且对体细胞和生殖系谱系的贡献能力保留程度不同。