大鼠脂肪干细胞诱导培养成为神经细胞并鉴定的实验研究  

Experimental study on culture of nerve cells via induction of rat adipose derived stem cells and its identification

作　　者：羊书勇[1] 杜敏 郑维银[1] 李晨军[1] 李焰[1] 蒋佶[1] 鄢兰元[1] 吴坡[1] 李浩[1] 

机构地区：[1]成都军区总医院附属口腔医院,成都610017 [2]南充市中心医院口腔科

出　　处：《西南国防医药》2016年第4期349-353,共5页Medical Journal of National Defending Forces in Southwest China

基　　金：四川省卫生厅课题资助(120568)

摘　　要：目的探究大鼠脂肪干细胞(r ADSCs)诱导成神经细胞的可行性。方法第一步,通过酶消化法获取r ADSCs并进行培养,通过CCK-8法检测细胞增殖情况,并应用免疫组化进行鉴定,确定获取的细胞为r ADSCs;第二步以原代培养的r ADSCs为研究对象,传代至第2代后,采用神经诱导培养基,诱导r ADSCs成神经细胞,诱导所得细胞通过形态学及免疫荧光进行鉴定。结果第一步中细胞培养增殖良好,增殖曲线显示细胞生长活跃,增殖能力强;免疫组化检测相关抗原CD90、CD105为阳性,与造血干细胞相关抗原CD45为阴性,说明所得细胞为r ADSCs。第二步中,诱导培养的细胞神经球免疫荧光染色Nestin阳性,细胞免疫荧光染色MAP-2阳性,结合形态学可确定ADSCs已成功诱导分化成神经细胞。结论 r ADSCs生长活跃,表现出稳定的增长及增殖能力,并可定向诱导分化为神经细胞。Objective To explore the feasibility of neural differentiation of rat adipose derived stem cells（r ADSCs） into nerve cells.Methods Firstly, r ADSCs were obtained by collagenase digestion and then cultured. The cell proliferation was detected by CCK-8.The cells were identified by immunohistochemical method in order to confirm they were r ADSCs. Secondly, primarily cultured r ADSCs were selected as research objects which were induced into nerve cells via neural induction culture medium after the second transfer of culture. And the induced cells were indentified by morphology and immunofluorescence. Results At the first step, the cell culture and proliferation were good. The growth curve showed the cell growth was active and the proliferation ability was strong.Immunohistochemistry detected that the related antigen CD90 and CD105 were positive; the hemopoietic stem cell related antigen CD45 was negative, which indicated that the obtained cells were r ADSCs. At the second step, the immunofluorescence staining Nestin of the cultured cellular neurosphere was positive, and the cellular immunofluorescence MAP-2 was positive. Those results combined with morphology could confirm that ADSCs were successfully induced and differentiated into nerve cells. Conclusion The r ADSCs growth is active and presents stable status and proliferation ability. Those cells can be induced and differentiated into nerve cells.

关 键 词：脂肪干细胞 诱导 神经细胞 大鼠 

分 类 号：R318[医药卫生—生物医学工程]