Chang I K, Yoshiki A, Kusakabe M, Tajima A, Chikamune T, Naito M, Ohno T
RIKEN Cell Bank, Institute of Physical and Chemical Research (RIKEN), Tsukuba Science City, Japan.
Cell Biol Int. 1995 Jul;19(7):569-76. doi: 10.1006/cbir.1995.1103.
Intrinsic primordial germ cells (PGCs) from stage 27 (5-day-old) chick embryonic germinal ridges were cultured in vitro for a further 5 days, and shown to proliferate on stroma cells derived from the germinal ridge. To determine whether these cultured PGCs could colonize and contribute to the germ-line, PGCs were isolated by gentle pipetting, labeled with PKH26 fluorescent dye and injected into the blood stream of stage 17 (2.5-day-old) chick embryos. The recipient embryos were incubated until they reached stage 28. Thin sections of these embryos were analysed by fluorescent confocal laser microscopy. These analyses showed that the labeled donor PGCs had migrated into the germinal ridges of the recipient embryos, and transplanted PGCs had undergone at least 3-7 divisions. These results suggest that PGCs that had passed far beyond the migration stage in vivo were still able to migrate, colonize and proliferate in recipient chick embryonic gonads.
取自第27期(5日龄)鸡胚生殖嵴的内源性原始生殖细胞(PGCs)在体外进一步培养5天,结果显示它们能在源自生殖嵴的基质细胞上增殖。为了确定这些培养的PGCs是否能够定殖并对种系产生贡献,通过轻柔吹打分离出PGCs,用PKH26荧光染料标记,然后注入第17期(2.5日龄)鸡胚的血流中。将受体胚胎孵化至第28期。通过荧光共聚焦激光显微镜对这些胚胎的薄切片进行分析。这些分析表明,标记的供体PGCs已迁移到受体胚胎的生殖嵴中,并且移植的PGCs至少经历了3 - 7次分裂。这些结果表明,在体内已远远超过迁移阶段的PGCs仍能够在受体鸡胚性腺中迁移、定殖和增殖。