兔骨髓基质干细胞的培养、鉴定及标记的实验研究  

Experimental research of culture,identification and mark of rabbit bone marrow stroma stem cells

作　　者：邓辰红 王飞 王文兰 朱华 

机构地区：[1]内蒙古自治区人民医院儿科,010020 [2]内蒙古自治区人民医院骨科

出　　处：《中国实用医药》2016年第32期194-196,共3页China Practical Medicine

基　　金：内蒙古自治区自然基金(项目编号:2012MS1157)

摘　　要：目的对兔骨髓基质干细胞的培养方法、鉴定及标记技术进行初步研究。方法采用贴壁法培养幼年兔骨髓细胞,经反复传代细胞逐渐纯化,以流式细胞术检测细胞表面抗原。以5-溴脱氧尿嘧啶核苷(Brd U)标记骨髓基质干细胞,将骨髓基质干细胞注入兔脑内,2周后取脑作石蜡切片,进行免疫荧光染色。结果通过贴壁法培养幼年兔骨髓细胞可获得较纯化的细胞,并能在体外长期培养。CD31的阴性率为5.67%,CD34的阴性率为4.31%,CD45的阴性率为4.42%,CD44的阳性率为98.82%,CD71的阳性率为99.17%,CD90的阳性率为98.23%。提示CD31、CD34、CD45的结果为呈阴性,CD44、CD71、CD90的结果为阳性,Brd U能够掺DNA合成期的骨髓基质干细胞。结论贴壁法可获得高纯度的骨髓基质干细胞,Brd U可作为骨髓基质干细胞体内示踪的理想标记物。Objective To briefly research culture, identification and mark of rabbit bone marrow stroma stem cells. Methods Bone marrow cells of young rabbits were cultured by adhesion method for purification by repeated passage, and cell surface antigen was detected by flow cytometry. Bone marrow stroma stem cells marked by 5-bromodeoxyuridine （BrdU） was injected into rabbit brain for immunofluorescent staining of brain paraffin section after 2 weeks. Results Bone marrow cells cultured by adhesion method provided purified ceils suitable for long-term culture in vitro. CD31 had negative rate as 5.67%, CD34 had negative rate as 4.31%, CD45 had negative rate as 4.42%, CD44 had positive rate as 98.82%. CD71 laad positive rate as 99.17%. CD90 had positive rate as 98.23%. CD31, CD34 and CD45 showed negative outcome, and CD44, CD71 and CD90 showed positive outcome. BrdU was suitable to be taken by bone marrow stroma stem ceils in DNA synthesis. Conclusion Adhesion method provides bone marrow stroma stem cells with high purification, and BrdU can be used as ideal marker for in vivo tracking bone marrow stroma stem cells.

关 键 词：骨髓基质干细胞 CD分子 5-溴脱氧尿嘧啶核苷 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]