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小鼠睾丸gdnf基因的克隆及其在支持细胞中的表达 被引量：2

Cloning of gdnf in the Mouse Testis and Its Expression in Sertoli Cells
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摘要 目的:从小鼠睾丸中克隆胶质细胞源神经营养因子基因gdnf,构建真核表达载体,并转染支持细胞,以便用作培养精原干细胞(SSCs)的滋养层。方法:以正常成年昆明鼠为材料,提取小鼠睾丸组织中总RNA后,以RT-PCR技术克隆小鼠睾丸gdnf基因,构建真核表达载体,并转染TM4细胞(睾丸支持细胞株),在转染后40h进行免疫荧光鉴定。结果:成功克隆小鼠睾丸gdnf基因的cDNA,测序正确,免疫荧光细胞染色显示转染后的支持细胞中有GDNF蛋白表达。结论:本研究为以转染了gdnf基因的支持细胞作饲养层培养SSCs奠定了基础。 Objective： To clone the glial cell line-derived neurotrophic factor （GDNF） from the mouse testis, construct the eukaryotic expression vector and transfect this vector into Sertoli cells in order to use the gdnf-transfected Sertoli cells as the feeder layer to cultivate spermatogonial stem cells （SSCs）. Methods ： Total RNA was extracted from the testes of normal mature mice and gdnf was cloned and amplified using RT-PCR, inserted into the eukaryotic expression vector and transfected into sertoli cells （TM4 cell line）. Immunofluorescence with anti-GDNF antibodies was performed at 40 h following the transfection. Results ： gdnf cDNA was cloned successfully, and GDNF expressed after transfected into Sertoli cells. Conclusion ： This study provides a basis for culturing SSCs with gdnf-transfected Sertoli cells as the feeder layer.
作者 段翠密 李恩中 张世庆 王常勇 李德雪
机构地区 军事医学科学院基础医学研究所
出处 《中华男科学杂志》 CAS CSCD 2007年第11期975-978,共4页 National Journal of Andrology
基金 国家自然科学基金资助(30471246)
关键词 睾丸 GDNF基因 转染 支持细胞 小鼠 testis gdnf gene transfection Sertoli cell mouse
分类号 Q78 [生物学—分子生物学]
  • 相关文献

参考文献23

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共引文献26

同被引文献11

  • 1李恩中,李德雪,张世庆,王常勇,张学明,王海滨,鲁景艳,王静.以支持细胞为饲养层培养小鼠精原干细胞[J].动物学报,2006,52(4):774-779. 被引量：17
  • 2张世庆,李德雪,李恩中,王常勇,鲁景艳.Bmi1基因研究进展[J].生物技术通讯,2007,18(2):274-276. 被引量：2
  • 3Kubota H, Avarbock MR, Brinster RL. Growth factors essential for self-renewal and expansion of mouse permatogonial stem cells[J]. Proc Natl Acad Sci USA, 2004, 101(47): 16489--16494.
  • 4Brinster RL. Germline stem cell transplantation and transgenesis[J]. Science, 2002, 296(5576): 2174--2176.
  • 5Kanatsu-Shinohara M, Takehashi M, Shinohara T. Brief History, Pitfalls, and Prospects of Mammalian Spermatogonial Stem Cell Research[J]. Cold Spring Harb Symp Quant Biol, 2008, 73: 33.
  • 6Kubota H, Avarboek MR, Brinster RL. Culture conditions and single growth factors affect fate determination of mouse spermatogonial stem cells[J]. Biol Reprod, 2004, 71(3): 722--731.
  • 7Meng X, Lindahl M, Hyvonen ME, et al. Regulation of cell fate decision of undifferentiated spermatogonia by GDNF[J]. Science, 2000, 287(5457): 1489--1493.
  • 8Brinster RL. Germline stem cell transplantation and transgenesis[J]. Science, 2002, 296: 2174--2176.
  • 9Nagano M, Ryu BY, Brinster C J, et al. Maintenance of mouse male germ line stem cells in vitro[J]. Biol Reprod, 2003, 68(6): 2207--2214.
  • 10Zhang Shiqing, Li Dexue, Li Enzhong, et al. Expression localization of Bmil in mice testis[J]. Mol Cell Endocrinol, 2008, 287(1-2): 47--56.

引证文献2

中华男科学杂志
2007年 第11期

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