Chicken stem cell factor enhances primordial germ cell proliferation cooperatively with fibroblast growth factor 2.
Chicken stem cell factor enhances primordial germ cell proliferation cooperatively with fibroblast growth factor 2.
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DOI：
10.1262/jrd.2015-128
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发表时间：
2016-04-22
期刊：
影响因子：
--
通讯作者：
Tagami T
中科院分区：
文献类型：
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作者：
Miyahara D;Oishi I;Makino R;Kurumisawa N;Nakaya R;Ono T;Kagami H;Tagami T
最近开发了鸡原始生殖细胞（PGC）的体外培养系统，但参与PGC增殖的生长因子在很大程度上是未知的。在本研究中，我们研究了鸡干细胞因子（chSCF）对鸡PGC体外增殖的影响。我们建立了两种饲养细胞系（水牛大鼠肝细胞；BRL 细胞），它们稳定表达假定的分泌形式的 chSCF (chSCF1-BRL) 和膜结合形式的 chSCF (chSCF2-BRL)。将培养的 PGC 系在含有成纤维细胞生长因子 2 (FGF2) 的 chSCF1 或 chSCF2-BRL 饲养细胞上孵育，并研究每种 chSCF 同工型的生长效应。在chSCF2-BRL细胞上培养的PGC在培养20天时的体外增殖率比在chSCF1-BRL细胞上培养的PGC高3倍以上，比在正常BRL细胞上培养的PGC高5倍以上。因此，使用chSCF2-BRL饲养层对于鸡PGC的体外增殖是有效的。然而，在没有 FGF2 的情况下，没有观察到 chSCF2-BRL 上 PGC 增殖的加速，这表明 chSCF2 将充当 FGF2 的增殖辅因子。我们将chSCF2-BRL细胞上培养的PGC转移到受体胚胎中，产生种系嵌合鸡，并通过后代测试评估培养的PGC的种系能力。获得供体后代，种系传播率为3.39%。本研究的结果表明，chSCF2 与 FGF2 配合，可在体外诱导保留种系能力的鸡 PGC 过度增殖。
An in vitro culture system of chicken primordial germ cells (PGCs) has been recently developed, but the growth factor involved in the proliferation of PGCs is largely unknown. In the present study, we investigated the growth effects of chicken stem cell factor (chSCF) on the in vitro proliferation of chicken PGCs. We established two feeder cell lines (buffalo rat liver cells; BRL cells) that stably express the putative secreted form of chSCF (chSCF1-BRL) and membrane bound form of chSCF (chSCF2-BRL). Cultured PGC lines were incubated on chSCF1 or chSCF2-BRL feeder cells with fibroblast growth factor 2 (FGF2), and growth effects of each chSCF isoform were investigated. The in vitro proliferation rate of the PGCs cultured on chSCF2-BRL at 20 days of culture was more than threefold higher than those cultured on chSCF1-BRL cells and more than fivefold higher than those cultured on normal BRL cells. Thus, use of chSCF2-BRL feeder layer was effective for in vitro proliferation of chicken PGCs. However, the acceleration of PGC proliferation on chSCF2-BRL was not observed without FGF2, suggesting that chSCF2 would act as a proliferation co-factor of FGF2. We transferred the PGCs cultured on chSCF2-BRL cells to recipient embryos, generated germline chimeric chickens and assessed the germline competency of cultured PGCs by progeny test. Donor-derived progenies were obtained, and the frequency of germline transmission was 3.39%. The results of this study demonstrate that chSCF2 induces hyperproliferation of chicken PGCs retaining germline competency in vitro in cooperation with FGF2.