氦-氖激光对大鼠骨髓间充质干细胞生长及神经分化潜能的作用

摘要

目的 观察氦-氖(He-Ne)激光治疗仪对大鼠骨髓间充质干细胞(BMSCs)生长和神经分化潜能的作用.方法 采用全骨髓培养法获取Sprague-Dawley大鼠的原代BMSCs,取第3代培养的细胞,分为实验组(He-Ne激光治疗仪处理10 min)和对照组(空气暴露相同时间).采用CCK8法检测细胞的增殖活性,流式细胞仪检测细胞凋亡,锥虫蓝染色测定细胞活力,并采用细胞免疫荧光技术鉴定BMSCs的神经干细胞分化潜能.结果 ①细胞呈纤维状贴壁生长,表型鉴定结果为CD90、CD29表达阳性,CD45表达阴性,符合BMSCs的表型特点.② 实验组和对照组的细胞增殖活性比较,差异无统计学意义,P>0.05.③ 实验组细胞的早期凋亡率为(1.10±0.26)%,与对照组细胞的(1.43±0.06)%比较,差异无统计学意义,P>0.05.实验组的细胞活力为(96.2±0.6)%,与对照组的(96.2±0.7)%比较,差异亦无统计学意义,P>0.05.④实验组和对照组的BMSCs在神经干细胞培养液中培养后,均可呈现悬浮生长的神经球;免疫荧光染色显示,神经球中细胞的Nestin染色阳性.结论 He-Ne激光治疗仪对BMSCs的增殖和活力无明显影响,未引起细胞凋亡,未改变BMSCs向神经干细胞分化的潜能.%Objective To observe the effects of He-Ne laser therapy instrument on bone marrow mesenchymal stem cell ( BMSC) growth and neural differentiation potential. Methods The primary BMSCs from Sprague-Dawley rats were Laken using whole bone marrow culture. The passage 3 cells were randomly divided into an experimental group ( He-Ne laser therapy instrument Irealed for 10 min) and a conlrol group ( exposing in the air for the same time) according to whether they were Ireated wilh the He-Ne laser therapy inslrumenl or nol. The cell proliferation aclivily was measured wilh cell counting kit-8 ( CCK-8 ) assay. Apoptosis detection was performed using flow cytometry. The cell vitality was detected by Trypan blue staining. Human neural stem cell differentiation potential was identified by immunofluorescence tech-nique. Results ①Cells showed a fibroblast-like adherent growth. The phenotypic identification results revealed thai the expressions of CD29 and CD90 were positive, but the expression of CD45 was negative, and it is consistent with the characteristics of the BMSC phenotype. ②There was no significant difference in the proliferative activity between the experiment group and the controt group,P ＞ 0. 05. ③There were no significant differences between the early apoptosis rate (1. 10 ± 0. 26% ) in the experiment group and that (1.43±0.06%) in the control group, as well as the cell viability (96. 2 ± 0. 6% ) in experiment group and that (96.2 ±0.7%) in the control group. ④Similar io the control group, BMSCs in the experiment group showed the suspended neurospheres after being cultured in neural stem cell culture medium. Immu-nofluorescence staining showed that the nestin slaining of neurosphere cetts was positive. Conclusion He-Ne laser therapy instrument has no significant effects on the proliferation and viability of BMSCs. It does not cause apoptosis and change the potential of BMSCs to neural stem cell differentiation.