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鸡胚胎原始生殖细胞体外培养

Culture of Chicken Embryonic Primordial Germ Cells in Vitro

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【作者】 谢蓓; 黄海; 胡小芬; 曹祥荣; 黄启忠; 李震;

【Author】 Bei Xie1,2, Hai Huang1,2, Xiao-Fen Hu2,4, Xiang-Rong Cao1, Qi-Zhong Huang2, Zhen Li2,3* ( 1 School of Life Science, Nanjing Normal University, Nanjin 210097, China; 2 Division of Animal Genetic Engineering, Shanghai Municipal Key Laboratory of Agri-genetics and Breeding, Shanghai 201106, China; 3 Animal Husbandry and Veterinary Research Institute, Shanghai Academy of Agricultural Sciences , Shanghai 201106, China; 4Jiangxi Provincial Key Laboratory for Animal Biotechnology, Jiangxi Agricultural University, Nanchan 330045, China )

【机构】 南京师范大学生命科学学院; 上海市农业遗传育种重点实验室; 上海市农业遗传育种重点实验室 南京210097上海市农业遗传育种重点实验室; 上海201106; 南京210097上海市农业遗传育种重点实验室; 上海201106江西农业大学江西省动物生物技术重点开放实验室; 南昌330045; 南京210097; 上海201106上海市农业科学院畜牧兽医研究所; 上海201106;

【摘要】 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。

【Abstract】 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 .

【基金】 上海市农业科学院中青年学术技术带头人培养基金~~
  • 【文献出处】 细胞生物学杂志 ,Chinese Journal of Coll Biology , 编辑部邮箱 ,2005年02期
  • 【分类号】S814.8
  • 【被引频次】14
  • 【下载频次】224
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