Production of quail (Coturnix japonica) germline chimeras derived from in vitro-cultured gonadal primordial germ cells
Production of quail (Coturnix japonica) germline chimeras derived from in vitro-cultured gonadal primordial germ cells
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DOI：
10.1002/mrd.20821
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发表时间：
2008-02-01
影响因子：
2.5
通讯作者：
Han, Jae Yong
中科院分区：
文献类型：
--
作者：
Park, Tae Sub;Kim, Mi A.;Han, Jae Yong
本实验室先前的一份报告记录了通过转移性腺原始生殖细胞（gPGCs）成功地生产鹌鹑（Cotumix japonica）生殖系嵌合体。随后，本研究旨在评估gPGC是否可以在体外维持较长时间，并且这些培养的PGC在移植到受体胚胎中后是否可以诱导生殖系传递。在实验1中，将来自两个品系（野生型羽毛（WP）和黑（D）鹌鹑）的性腺细胞体外培养10天。使用抗体QCR 1，我们检测到WP数量的连续、显著（P=0.0002）增加，但未检测到D、PGC。QCR1阳性WP菌落在培养7天后开始形成。在培养的第10天，803 WP PGC作为一个连续增加的结果，而没有D PGC集落可以检测到和D性腺基质细胞卷起。这可能与两种品系的原始生殖细胞或性腺基质细胞的差异有关。在实验2中，WP PGC集落在体外培养20天，并将培养10或20天的PGC显微注射到181个受体D胚胎的背侧腹。移植胚胎孵化率为19.3%，其中25只（71.4%）已达性成熟。性成熟的幼体的测交，分别导致3（10天，33.3%）和8（20天，50.0%）的种系嵌合体。这份报告是第一次描述成功生产的生殖嵌合体的转移，在体外培养的gPGCs在鹌鹑。
A previous report from our laboratory documented successful production of quail (Cotumix japonica) germline chimeras by transfer of gonadal primordial germ cells (gPGCs). Subsequently, this study was designed to evaluate whether gPGCs can be maintained in vitro for extended period, and furthermore, these cultured PGCs can induce germline transmission after transfer into recipient embryos. In experiment 1, gonadal cells from the two strains (wild-type plumage (WP) and black (D) quail) were cultured in vitro for 10 days. Using antibody QCR1, we detected a continuous, significant (P=0.0002) increase in the number of WP, but not D, PGCs. QCR1-positive WP colonies began to form after 7 days in culture. On Day 10 of culture, 803 WP PGCs were present as a result of a continuous increase, whereas no D PGC colonies could be detected and the D gonadal stroma cells were rolled up. Differences in the PGCs or the gonadal stroma cells of the two different strains might account for these differences. In experiment 2, WP PGC colonies were maintained in vitro up to Day 20 of culture, and 10- or 20-day-cultured PGCs were microinjected into dorsal aortas of 181 recipient D embryos. Thirty-five (19.3%) of the transplanted embryos hatched after incubation, and 25 (71.4%) of the hatchlings reached sexual maturity. Testcrossing of the sexually mature hatchlings; resulted in three (10 days, 33.3%) and eight (20 days, 50.0%) germline chimeras respectively. This report is the first to describe successful production of germline chimera by transfer of in vitro-cultured gPGCs in quail.