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目的:研究去分化间充质干细胞(de-differentiated mesenchymal stem cells,De-MSCs)成骨再分化能力。方法:以MSCs为对照,分别采用Cell Count Kit(CCK8)法、实时定量PCR(qPCR)、碱性磷酸酶(ALP)活性和茜素红染色法检测De-MSCs增殖能力、成骨相关基因、成骨再分化能力等。结果:De-MSCs可表达干细胞表面标志,在成骨培养基中增殖能力高于MSCs(P<0.05),成骨相关基因(Runx2、Osterix、Bmp2)表达明显增加(P<0.05),ALP活性检测发现De-MSCs明显高于MSCs组(P<0.05),28 d后茜素红染色可见红色结节。结论:De-MSCs具备某些MSCs特征,但是体外再次成骨效率明显提高,De-MSCs可作为更优质种子细胞,为再生医学开拓新的思路。
Objective: To study the osteogenic differentiation of de-differentiated mesenchymal stem cells (De-MSCs). Methods: The proliferation of De-MSCs was detected by Cell Count Kit (CCK8) assay, qPCR, Alkaline phosphatase (ALP) and alizarin red staining, respectively. Osteogenic differentiation and so on. Results: De-MSCs could express the surface markers of stem cells. The proliferation ability of MSCs in osteogenic medium was higher than that of MSCs (P <0.05), and the expression of osteogenesis related genes (Runx2, Osterix, Bmp2) De-MSCs were found to be significantly higher than MSCs group (P <0.05). After 28 days, red nodules were observed by alizarin red staining. Conclusion: De-MSCs possess some features of MSCs, but the efficiency of re-osteogenesis in vitro is significantly improved. De-MSCs can be used as more excellent seed cells, opening up new ideas for regenerative medicine.