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目的:比较骨髓间充质干细胞(BMSCs)和脂肪间充质干细胞(ADSCs)成骨分化时序性特点及其潜能的差异。方法:分离提取同一SD大鼠的BMSCs和ADSCs两种细胞进行培养,通过免疫荧光染色和Von Kossa染色分别观察两种细胞经成骨诱导培养后的成骨分化蛋白I型胶原(Col1α1)的分泌和矿化结节沉积。利用实时荧光定量PCR(RT-q PCR)方法检测两种细胞成骨分化相关基因的表达情况。结果:免疫荧光染色结果显示第7 d时ADSCs的Col1α1表达强度较BMSCs弱,但在14 d和21 d时两者差异并不明显。Von Kossa染色显示21 d时两者均呈现出较好的矿化能力。RT-q PCR结果显示ADSCs成骨相关基因表达水平在早中期(7、14 d)低于BMSCs(P<0.05),而在晚期(21、28 d)无显著性差异(P>0.05)。结论:ADSCs在早期成骨分化水平低于BMSCs,但在晚期接近BMSCs,显示出良好的成骨分化潜能,在骨组织工程中将有广泛的应用前景。
OBJECTIVE: To compare the osteogenic differentiation characteristics of bone marrow mesenchymal stem cells (BMSCs) and adipose tissue-derived mesenchymal stem cells (ADSCs) and their potential differences. Methods: BMSCs and ADSCs were isolated and extracted from the same Sprague-Dawley rats. The immunofluorescence staining and Von Kossa staining were used to observe the secretion of osteogenic differentiation type I collagen (Col1α1) And mineralized nodular deposits. Real-time quantitative PCR (RT-q PCR) was used to detect the expression of osteogenic differentiation-related genes in both cells. Results: Immunofluorescence staining showed that the expression of Col1α1 in ADSCs was weaker than that in BMSCs on the 7th day, but the difference was not obvious on the 14th and 21st days. Von Kossa staining showed that both showed good mineralization on the 21st day. The results of RT-q PCR showed that the expression of osteogenesis-related genes in ADSCs was lower than that in BMSCs at early and middle stages (P <0.05), but not in late stage at 21 and 28 days (P> 0.05). CONCLUSION: ADSCs have a lower level of early osteogenic differentiation than BMSCs, but close to BMSCs in late stage, indicating good osteogenic differentiation potential. ADSCs will have a wide range of applications in bone tissue engineering.