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目的:探讨骨髓间充质干细胞(bone marrow mesenchymal stem cells,BMMSCs)膜片的生物学性能以及骨形成蛋白7(bone morphogenetic protein 7,BMP-7)对BMMSCs膜片成骨分化的影响。方法:分离培养SD大鼠原代BMMSCs,采用维生素C诱导构建大鼠BMMSCs膜片,实验分为自然分化组、添加BMP-7的骨诱导组以及标准骨诱导组,采用HE染色观察,CCK-8检测膜片细胞增殖情况以及RT-PCR检测细胞膜片成骨分化水平。结果:分离大鼠BMMSCs诱导10~14 d后获得白色膜状细胞膜片。镜下观察细胞呈梭形重叠生长,细胞间紧密连接并有大量细胞外基质(ECM)分泌。CCK-8显示膜片细胞密度随时间递增。与BMMSCs膜片相比,BMP-7诱导的细胞膜片高表达ALP、Osteocalcin等成骨基因。结论:BMP-7诱导的大鼠BMMSCs细胞膜片成骨分化能力强,在骨组织工程中优势显著。
Objective: To investigate the biological properties of bone marrow mesenchymal stem cells (BMMSCs) and the effect of bone morphogenetic protein 7 (BMP-7) on the osteogenic differentiation of BMMSCs. Methods: The primary BMMSCs of SD rats were isolated and cultured, and the membrane of rat BMMSCs was induced by vitamin C. The experiment was divided into natural differentiation group, osteoinductive group with BMP-7 addition and standard osteoinductive group. HE staining, CCK- 8 detection of membrane cell proliferation and RT-PCR detection of cell membrane osteogenic differentiation. Results: The white membrane-like membrane was obtained after 10 ~ 14 d of rat BMMSCs isolation. Microscopically observed cells were spindle-shaped overlapping growth, tight junctions between cells and a large number of extracellular matrix (ECM) secretion. CCK-8 shows patch cell density increases with time. Compared with BMMSCs, BMP-7 induced high expression of osteoblast genes, such as ALP and Osteocalcin. CONCLUSION: BMP-7 induced osteogenic differentiation of rat BMMSCs is superior in bone tissue engineering.