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目的:观察不同培养液及不同培养时期等因素对体外培养人脑胶质瘤细胞CD133阳性(CD133~+)细胞比率的影响。方法:用干细胞培养液(含EGF、FGF-2、B27)培养6例原代胶质瘤细胞和3例对照细胞株,用流式细胞仪检测培养第0、3、7、28、60、90和120天时不同细胞的CD133~+细胞比率;免疫组化观察胶质瘤干细胞的多向分化能力;用血清、无血清和干细胞3种培养液分别培养2例长期培养的胶质瘤SHG62、SHG66细胞系以及对照的U87和U251细胞株,观察胶质瘤中CD133~+细胞在不同培养液中的相互转换。结果:胶质瘤CD133~+细胞比率随培养时间推移明显下降(P<0.05),1周左右均降至<1%的低水平,而细胞株则比较稳定;干细胞培养液培养的细胞中CD133~+细胞比率明显高于血清和无血清培养液培养(P<0.01);此外,胶质瘤细胞中的干细胞具有多向分化的能力,并且可以在干细胞和血清培养液中互相转换。结论:原代胶质瘤体外培养过程中CD133~+细胞比率逐渐下降,而干细胞培养液能明显提高CD133~+细胞比率,并且CD133~+细胞可以在不同培养液中相互转换。说明生长环境对干细胞比率影响较大,提示可能存在尚未明了的影响干细胞生长分化的相关因子。
Objective: To observe the effects of different culture media and different culture periods on the ratio of CD133 positive cells (CD133 ~ +) in cultured human glioma cells. Methods: Six primary glioma cells and three control cells were cultured in stem cell culture medium (containing EGF, FGF-2 and B27). Flow cytometry was used to detect the proliferation of primary glioma cells (0,3,7,28,60, The percentage of CD133 ~ + cells in different cells at 90 and 120 days was observed by immunohistochemistry. The long-term cultured glioma SHG62 was cultured in serum, serum-free and stem-cell-free medium. SHG66 cell line and control U87 and U251 cell lines were observed in glioma CD133 ~ + cells in different culture medium conversion. Results: The percentage of CD133 + cells in glioma cells decreased significantly with time (P <0.05), but decreased to <1% at 1 week, while cell lines were stable. CD133 ~ + Cells were significantly higher than those in serum and serum-free medium (P <0.01). In addition, stem cells in glioma cells were capable of multi-directional differentiation and could convert to each other in stem cells and serum medium. CONCLUSION: The ratio of CD133 ~ + cells in primary glioma cells decreased gradually in vitro, while the percentage of CD133 ~ + cells in stem cells could be significantly increased. CD133 ~ + cells could be transformed in different culture media. Indicating that the growth environment on the stem cell ratio greater impact, suggesting that there may be unknown factors that affect stem cell growth and differentiation.