接种密度对大鼠髓核间充质干细胞的影响

发布时间：2018-01-19 10:08

本文关键词： 髓核间充质干细胞低密度接种传代免疫表型大鼠　出处：《中国矫形外科杂志》2017年11期 　论文类型：期刊论文

【摘要】：[目的]比较不同细胞接种密度传代培养大鼠髓核间充质干细胞(nucleus pulposus-derived mesenchymal stem cells,NPMSCs)对NPMSCs的形态及表面标志物的影响。[方法]取12周龄SD大鼠椎间盘髓核组织体外消化获得原代细胞,待达到60%~70%融合时消化,并按接种密度分为三组,分别为低密度组(5个细胞/cm~2)、中密度组(100个细胞/cm~2)、正常密度组(10 000个细胞/cm~2)。培养至达到传代条件后,各组细胞均按10 000个细胞/cm~2在体外进行传代接种,进行形态学观察并检测干细胞的特征性表面标志(CD29、CD34、CD44、CD45、CD90)的表达情况。[结果]在细胞形态学方面,各接种密度组细胞均可呈旋涡状贴壁生长,但低密度组以集落样生长,细胞多呈长梭形,形态一致;中密度组和高密度组细胞中类圆形、多边形细胞增多,大量细胞出现形态不规则、细胞宽大扁平、折光性差。在细胞表面标志物方面,低密度组CD44、CD90的表达率明显高于中密度组和高密度组(P0.05),而CD29、CD34、CD45差异无统计学意义(P0.05)。[结论]低密度接种法可提高大鼠NPMSCs CD44及CD90的阳性率,纯化NPMSCs。
[Abstract]:[Objective] to compare the subculture of rat nucleus pulposus mesenchymal stem cells (MSCs) with different cell densities. Nucleus pulposus-derived mesenchymal stem cells. The effect of NPMSCs on the morphology and surface markers of NPMSCs. [Methods the primary cells were digested from the nucleus pulposus of SD rats at 12 weeks of age in vitro and digested by 70% fusion and divided into three groups according to the density of inoculation. They were divided into low density group (5 cells / cm ~ (2)), medium density group (100 cells / cm ~ (2)) and normal density group (10 000 cells / cm ~ (2)). All the cells in each group were inoculated in vitro according to 10 000 cells / cm ~ (2) to observe the morphology and to detect the characteristic surface marker of stem cells: CD29, CD34 and CD44. Expression of CD45 + CD90). [Results: in the aspect of cell morphology, the cells in each inoculation density group all showed vortex adherent growth, but in the low density group, the cells grew like colony, and most of the cells were spindle-shaped, and the morphology was the same. In the medium density group and high density group, the cells were round, polygonal cells increased, a large number of cells appeared irregular, the cells were broad and flat, and the refractive index was poor. In terms of cell surface markers, low density group CD44. The expression rate of CD90 was significantly higher than that of middle density group and high density group, but the difference of CD29 CD34 and CD45 was not statistically significant. [Conclusion: low density inoculation can increase the positive rate of NPMSCs CD44 and CD90 in rats.
【作者单位】：南方医科大学第三临床医学院;海军总医院骨科;解放军第306医院航天城临床部;
【基金】：国家自然科学基金资助项目(编号:81470102;81171740)
【分类号】：R681.5
【正文快照】： 椎间盘退变是譬如椎间盘突出等脊柱退行性疾病的病理学基础。但是目前仍不明确椎间盘退变的相关机制。近年来,在机体的大多数组织及器官中,均发现了维持组织内稳态的重要组成部分——内源性干细胞,目前认为这些干细胞是组织或器官是否产生疾病的重要原因。最近,髓核组织中也发

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