鸡胚胎原始生殖细胞体外培养

Culture of Chicken Embryonic Primordial Germ Cells in Vitro

以14￣15期鸡胚血液为材料,采用Ficoll密度梯度离心方法,提取鸡胚胎原始生殖细胞(primordial germ cells,PGCs),在无基质细胞和基质细胞上分别进行体外培养。从实验结果可以看出:在含有胎牛血清(fetal bovine serum,FBS)、鸡血清(chicken serum,CS)、碱性成纤维细胞生长因子(bFGF)、人胰岛素样生长因子(hIGF-1)、小鼠白血病抑制因子(mLIF)和青、链霉素双抗的M199培养液中培养时,鸡PGCs最多能够存活4天;当采用细胞因子和5天鸡胚胎性腺基质细胞共培养时能存活23代且每代细胞增殖可达近10倍。提纯后的PGCs细胞冻存复苏后,经台盼蓝染色鉴定存活率可达80%左右。

Primordial germ cells (PGCs) were isolated from the blood of stage 14-15 chick embryos by using Ficoll density centrifugation. After isolated, primordial germ cells were cultured with or without stroma cells. The PGCs could survive 4 days when cultured in Medium 199 supplemented with FBS, chicken serum, bFGF, human IGF-1 and murine LIF, while they could be cultured for twenty-three generations when cultured on different growth factors and stromal cells derived from 5-day-old chicken germinal ridge. The PGCs could maintain a viabil- ity rate of 80% after being frozen and thawed .