新生牛雄性生殖细胞源类ES的培养及检测  

Culture and Cytochemical Analysis of ES-like Cells from Neonatal Calf Male Germ Stem Cells

作　　者：董武子[1] 沈文正[1] 华进联[1] 窦忠英[1] 

机构地区：[1]西北农林科技大学国家干细胞工程技术研究中心陕西分中心,杨凌712100

出　　处：《畜牧兽医学报》2007年第2期144-148,共5页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：国家自然科学基金项目(39970363);国家重点基础研究发展规划项目(G1999054300);西北农林科技大学青年专项基金(08080258)

摘　　要：采用贴壁速率差法分离纯化的新生牛雄性生殖干细胞（Male germ stem cell，mGSCs）和支持细胞（Calf sertoli cell，CSCs）；CSCs饲养层细胞一般维持时间不超过8d；新生牛雄性生殖干细胞在CSCs饲养层上1代培养至5～6d可形成类ES细胞集落，采用机械离散集落传代法有利于集落形成或存在，传代培养至少在前2代细胞集落存在；部分典型细胞集落的中心细胞AKP强阳性，周边细胞AKP弱阳性或阴性；免疫组化检测显示，细胞集落SSEA1强阳性、SSEA3弱阳性和Oct-4呈现阳性染色。表明mGSCs具有ES细胞的某些细胞特性。Neonatal calf male germ stem cells （mGSCs） and calf sertoli cells（CSCs） were separated and purified through the method of adhering velocity difference, and in vitro cultivation of mGSCs was studied on feeder layer cells of CSCs. The feeder layer cells of CSCs survived time did not exceed 8 d. ES-like clusters formed when mGSCs was cultured on CSCs till 5-6 d. The method of mechanical dispersion could favor formation and presence of clusters. At lest in first 2 passages, this clusters existed. Cells of part clusters center were AKP positive staining and cells of clusters margin were AKP light positive or negative staining. Immunohistochemical analysis of typical clusters showed SSEA1 strongly positive, SSEA3 light positive and Oct-4 positive. The results suggested neonatal calf mGSCs can form ES cells.

关 键 词：新生牛雄性生殖干细胞 新生牛支持细胞 类ES细胞 

分 类 号：Q813.1[生物学—生物工程]