两种方法培养骨质疏松症小鼠脂肪干细胞的比较

发布时间：2018-05-07 08:36

本文选题：干细胞 + 骨质疏松　；参考：《中国组织工程研究》2017年29期

【摘要】：背景:获得充足的脂肪干细胞是自体移植修复骨质疏松症骨缺损的前提,因此如何高效、简便、经济的获得自体原代脂肪干细胞是治疗骨质疏松症的关键。目的:采用组织块法和胶原酶消化法体外培养骨质疏松症小鼠脂肪干细胞,并对两种培养方法进行比较分析。方法:切除雌性C57BL/6小鼠双侧卵巢,制作骨质疏松模型,造模成功后取小鼠皮下脂肪组织,分别采用组织块法和胶原酶消化法分离培养脂肪干细胞。应用流式细胞仪检测第3代脂肪干细胞表面特异性抗原;培养7 d后,检测细胞产量;检测第3代脂肪干细胞的增殖;检测第4代脂肪干细胞的成脂及成骨分化能力。结果与结论:①组织块法培养第3代细胞CD34、CD146、Sca-1表达率分别为(15.22±1.85)%、(75.55±3.36)%、(83.48±4.22)%,胶原酶消化法第3代细胞CD34、CD146、Sca-1表达率分别为(13.46±2.21)%、(76.62±2.47)%、(84.84±3.56)%;②组织块法每毫克脂肪组织获得的细胞产量高于胶原酶消化法(P0.05);③培养24 h后,组织块培养法的细胞增殖率高于胶原酶消化法;④两种方法得到的脂肪干细胞均可向脂肪细胞和成骨细胞方向分化,两组间成脂量及成骨量比较差异无显著性意义(P0.05);⑤结果表明,组织块法比胶原酶消化法更适合体外培养脂肪干细胞,可为骨质疏松症患者自体移植脂肪干细胞治疗骨质疏松性骨折和骨缺损研究提供充足的细胞来源。
[Abstract]:Background: obtaining sufficient adipose stem cells is the premise of autotransplantation to repair bone defect of osteoporosis. Therefore, how to obtain autologous primary adipose stem cells efficiently, conveniently and economically is the key to the treatment of osteoporosis. Aim: to culture adipose stem cells of osteoporosis mice in vitro by tissue mass method and collagenase digestion method. Methods: the ovarium of female C57BL/6 mice was excised and the model of osteoporosis was made. The adipose tissue was isolated and cultured by tissue block method and collagenase digestion method. The surface specific antigen of the third generation of adipose stem cells was detected by flow cytometry, the cell production was detected after 7 days of culture, the proliferation of the third generation of adipose stem cells was detected, and the ability of adipogenic and osteogenic differentiation of the fourth generation of adipose stem cells was detected. Results and conclusion the expression rates of CD34, CD146, Sca-1 were 15.22 卤1.85, 75.55 卤3.36 and 83.48 卤4.22, respectively, and the expression rates of CD34 and CD146Sca-1 in the third passage were 13.46 卤2.21, 76.62 卤2.47 and 84.84 卤3.56%, respectively. The cell yield per milligram of adipose tissue by collagenase digestion was higher than that of collagenase elimination. After 24 h culture of P0. 05 and P0. 05, P0. 05 and P0. 05%. The proliferation rate of adipose stem cells obtained by tissue mass culture method was higher than that by collagenase digestion method. The adipocytes and osteoblasts were differentiated into adipocytes and osteoblasts. There was no significant difference in fat production and osteogenesis between the two groups. The results showed that the tissue block method was more suitable than collagenase digestion method to culture adipose stem cells in vitro. It can provide sufficient cell source for the treatment of osteoporotic fracture and bone defect by autologous transplantation of adipose stem cells in patients with osteoporosis.
【作者单位】：西南医科大学附属口腔医院口腔颌面外科;西南医科大学附属口腔医院口颌面修复重建和再生实验室;
【基金】：国家自然科学基金项目(81371125) 四川省科技厅项目(2014JY0044) 四川省教育厅项目(10ZB030) 四川省卫生厅项目(80170) 西南医科大学重点项目(201207) 2015年泸州市人民政府-西南医科大学科技战略合作项目[2015LZCYD-S05(2/12)] 西南医大口院[(2017)20号]~~
【分类号】：R580

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