论文部分内容阅读
次传代得到较纯的MSCs,倒置相差显微镜下观察细胞形态,应用免疫细胞化学方法对细胞的表面抗原标志进行检测。取3代MSCs,B-ME诱导6小时候后,倒置相差显微镜下观察细胞形态,应用免疫细胞化学方法鉴定诱导后细胞的表型特征。结果:倒置相差显微镜观察发现,接种后24h,细胞开始贴壁。培养3代以后,细胞呈梭形或多角形,折光性好,核为圆形、居中。免疫细胞化学检测细胞表面抗原显示:CD34-、CD45-、CD29+、CD44+、CD90+。诱导后细胞发出突起,逐渐生长延伸并彼此相连,形态学上具有神经细胞的明显特征。免疫细胞化学显示诱导后细胞NSE(神经元特异性烯醇化酶)阳性。结论:所采用的细胞分离培养方法简便可行,所获得的细胞其表型特征与文献报道一致,且具有向神经细胞分化的潜能.
Sub-passage to get more pure MSCs, inverted phase contrast microscope to observe the cell morphology, the application of immunocytochemistry to detect the cell surface antigenic markers. Three generations of MSCs were harvested. After 6 hours of B-ME induction, morphological changes were observed under inverted phase contrast microscope. Immunocytochemistry was used to identify the phenotypic characteristics of the induced cells. Results: Inverted phase contrast microscopy showed that after 24h of inoculation cells began to adhere. After three generations of culture, the cells are fusiform or polygonal, with good refractivity and round nuclei. Immunocytochemistry detection of cell surface antigen showed: CD34-, CD45-, CD29 +, CD44 +, CD90 +. After induction, the cells give rise to protrusions, grow gradually and are connected to each other, and have morphologically distinct features of nerve cells. Immunocytochemistry showed positive for cell NSE (neuron-specific enolase) after induction. Conclusion: The method of cell isolation and culture is simple and feasible. The phenotypes of the obtained cells are consistent with those reported in the literature, and have the potential to differentiate into nerve cells.