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目的:探讨模拟微重力(SMG)对骨髓间充质干细胞(MSCs)的增殖及向脂肪方向分化能力的影响。方法:第一部分将第三代的MSCs分为两组,分别在正常重力下(NG组)及微重力下(SMG组,采用回转模拟装置以30r/min回转模拟微重力),培养72h后,采用BrdU标记法检测两组细胞的增殖情况,细胞计数法绘制细胞生长曲线。Western Blot检测干细胞标志物Oct4、SSEA4的表达情况,第二部分将第三代MSCs分为三组:第一组在NG条件下培养后,加入脂肪方向诱导剂在NG条件下诱导、第二组在SMG条件下培养,在NG条件下诱导,第三组在SMG条件下培养,在SMG条件下诱导。7天后,油红O染色观察脂肪方向的诱导率,Western Blot检测过氧化物酶增殖物激活受体γ2(PPARγ2)以及Oct4的表达。结果:第一部分:流式细胞仪检测SMG组BrdU标记阳性率明显高于NG组,表明细胞增殖较快,Western Blot结果显示SMG组细胞中Oct4、SSEA4的表达量明显高于NG组,有统计学意义。第二部分:脂肪方向诱导后第一组细胞油红O染色阳性,Western Blot显示PPARγ2呈阳性表达,Oct4仅有微量表达,第二组油红O染色阳性表达率明显高于第一组,且PPARγ2表达较第一组增多,几乎未见Oct4的表达,第三组细胞油红O染色阴性,且几乎不表达PPARγ2,而Oct4表达较前两组升高。结论:模拟微重力可促进骨髓间充质干细胞增殖,提高其向脂肪方向分化的能力可能与微重力保持其未分化状态相关。
Objective: To investigate the effects of simulated microgravity (SMG) on the proliferation and adipogenic differentiation of mesenchymal stem cells (MSCs). Methods: The first part of the third generation of MSCs were divided into two groups, respectively, under normal gravity (NG group) and microgravity (SMG group, rotary simulation device to 30g / min rotary simulation of microgravity), after 72h, BrdU labeling method was used to detect the proliferation of two groups of cells, cell counting method to draw cell growth curve. Western Blot detection of stem cell markers Oct4, SSEA4 expression, the second part of the third generation of MSCs are divided into three groups: the first group cultured under NG conditions, the addition of fat direction inducer induced under NG conditions, the second group Cultured under SMG conditions, induced under NG conditions, the third group cultured under SMG conditions, and induced under SMG conditions. Seven days later, the induction rate of adipose tissue was observed by oil red O staining, and the expression of peroxisome proliferator-activated receptor γ2 (PPARγ2) and Oct4 were detected by Western Blot. Results: The first part: The positive rate of BrdU labeling in SMG group by flow cytometry was significantly higher than that in NG group, indicating that the cell proliferation was rapid. Western Blot showed that the expression of Oct4 and SSEA4 in SMG group was significantly higher than that in NG group Significance of learning. In the second part, the first group of cells were oily red O staining after induction of adipose tissue, the expression of PPARγ2 was positive by Western Blot, the expression of Oct4 was slight, the second group was significantly higher than that of the first group PPARγ2 expression increased compared with the first group, almost no expression of Oct4, the third group of cells oily red O staining, and almost no expression of PPARγ2, while Oct4 expression increased compared with the previous two groups. CONCLUSION: Simulated microgravity can promote the proliferation of bone marrow mesenchymal stem cells and enhance their ability to differentiate into adipose tissue, which may be related to the microgravity maintaining its undifferentiated state.