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目的:探索骨质疏松大鼠(OVX)骨髓间充质干细胞(BMSCs)的培养方法,进行鉴定并观察其生物学特性。方法:通过去卵巢法构建SD大鼠骨质疏松模型,在OVX组大鼠和假手术组(Sham)大鼠,分别冲取不同部位(肱骨、股骨和胫骨)的骨髓,贴壁法分离纯化BMSCs,并通过MTT法检测其增殖活性,流式细胞仪检测其表面抗原;对BMSCs进行成骨、成脂诱导,并与Sham组BMSCs对比,观察OVX组BMSCs成骨、成脂肪分化的差别。结果:在OVX大鼠,无法培养出股骨中的BMSCs,可培养出胫骨和肱骨的BMSCs,但前者细胞量少,后者量多;与Sham组相比较,成骨诱导后OVX组BMSCs的碱性磷酸酶活性和骨钙素分泌均较低;成脂诱导后,OVX组BMSCs生成的脂滴数目较多。结论:通过选取肱骨骨髓,加大细胞培养密度,可以成功培养出OVX大鼠的BMSCs,细胞量明显多于其他部位骨髓,可满足实验的样本量要求;大鼠BMSCs(OVX)具有增殖速率低、成骨能力弱、成脂能力强的生物学特性。
Objective: To explore the culture method of bone marrow mesenchymal stem cells (BMSCs) of osteoporosis rats (OVX), to identify and observe its biological characteristics. Methods: Osteoporosis model of SD rats was established by ovariectomy. Bone marrow of different parts (humerus, femur and tibia) were implanted into OVX group and sham group respectively. BMSCs were detected by MTT assay. The surface antigens were detected by flow cytometry. BMSCs were induced to osteoblast and adipogenesis, and compared with BMSCs in Sham group. The differences of osteogenic and adipogenic differentiation of BMSCs in OVX group were observed. RESULTS: In OVX rats, BMSCs in the femur could not be cultured and BMSCs in the tibia and humerus could be cultured, but the amount of the former was small and the latter was large. Compared with Sham group, the BMSCs in OVX group The activity of phosphatase and the secretion of osteocalcin were lower. After adipogenic induction, the number of lipid droplets generated by BMSCs in OVX group was higher. CONCLUSIONS: BMSCs of OVX rats can be successfully cultured by selecting the humerus bone marrow and increasing the cell culture density. The amount of BMSCs in OVX rats is obviously higher than that in other parts of bone marrow, which can meet the experimental requirements. BMSCs (OVX) , Osteogenic ability is weak, adipogenic ability of biological characteristics.