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

发布时间：2018-11-15 13:17

【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。
[Abstract]:Objective to explore the feasibility of (r ADSCs) induction of rat adipose stem cells into neural cells. Methods in the first step, the r ADSCs was obtained by enzyme digestion and cultured. The proliferation of the cells was detected by CCK-8 method and identified by immunohistochemistry. The obtained cells were identified as r ADSCs;. In the second step, the primary cultured r ADSCs was used as the research object. After the passage to the second generation, the neurons of r ADSCs were induced by nerve induction medium. The induced cells were identified by morphology and immunofluorescence. Results in the first step, the cells were cultured and proliferated well, and the proliferation curve showed that the cells were active in growth and strong in proliferative ability, and that the relative antigen CD90,CD105 was positive and the CD45 associated with hematopoietic stem cells was negative, indicating that the cells obtained were r ADSCs.. In the second step, the cultured cells were stained with Nestin and MAP-2 with immunocytofluorescence staining. It was confirmed by morphology that ADSCs had been successfully induced to differentiate into nerve cells. Conclusion r ADSCs is active in growth, stable in growth and proliferation, and can be induced to differentiate into nerve cells.
【作者单位】：成都军区总医院附属口腔医院;南充市中心医院口腔科;
【基金】：四川省卫生厅课题资助(120568)
【分类号】：R329.2

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