去卵巢对SD大鼠MSCs生物学特性的影响

目的:通过比较正常与双侧卵巢切除法复制的骨质疏松症(OP)模型大鼠骨髓间充质干细胞(MSCs)的形态学、生长曲线及细胞周期的差异,观察去卵巢对SD大鼠MSCs生物学特性的影响。方法:20只10月龄SD大鼠随机分成模型组和假手术组,每组10只;以双侧卵巢切除法复制OP模型;造模12周后,采用全骨髓贴壁法提取正常和OP大鼠MSCs;差速贴壁原理纯化MSCs;分别采用倒置相差显微镜、透射电子显微镜(TEM)、扫描电子显微镜(SEM)观察并比较正常与OP大鼠MSCs的形态学差异;MTT法检测正常与OP大鼠MSCs生长曲线;流式细胞仪检测正常与OP大鼠MSCs细胞周期。结果:OP大鼠MSCs细胞形态变得宽大、扁平,细胞内部细胞器减少,细胞活力下降,并有向脂肪细胞分化的现象发生;与正常组比较,OP大鼠细胞的增殖能力下降。结论:去卵巢导致大鼠MSCs形态变得宽大、扁平,体外增殖能力下降。

Effect of ovariectomy on biological characteristics of MSCs from SD rats

AIM: To investigate the effects of ovariectomy on the biological characteristics of the mesenchymal stem cells (MSCs) derived from the osteoporotic (OP) SD rats and normal rats by comparing the morphology, growth curve and cell cycle. METHODS: Female SD rats of 10-month old were randomly divided into model group and sham operation group (10 rats in each group). The osteoporotic model was established by ovariectomy. Twelve weeks after ovariectomy, the MSCs were isolated from normal and OP rats, and purified by differential time adherent method. The morphological difference of the MSCs between normal and OP rats was observed under inverted phase contrast microscope, transmission electronic microscope (TEM) and scanning electronic microscope (SEM). The growth curves of the MSCs were detected by MTT method. The cell cycles of the MSCs were determined by flow cytometry. RESULTS: Compared to the MSCs from normal rats, the morphology of the MSCs form OP rats became wider and flatter, the cell organelles were decreased and the cell viability was descended. The differentiation of the MSCs from OP rats into adipocytes appeared and the cell multiplication capacity was declined. CONCLUSION: The cell morphology of the MSCs derived from rats with ovariectomy changes obviously and the cell multiplication capacity is decreased.