兔下颌骨来源骨髓间充质干细胞体外分离培养及生物学行为的研究

发布时间：2018-06-18 09:51

本文选题：兔 + 下颌骨　；参考：《佳木斯大学》2012年硕士论文

【摘要】：目的：探讨兔下颌骨来源的骨髓间充质干细胞（bone marrow mesenchymal stemcells, BMMSCs）体外分离、培养及其生物学行为。方法：取兔下颌骨中的松质骨，密度梯度离心分离法收集单个核细胞，，贴壁培养，取第P2或P3代检测：（1）倒置显微镜下观察细胞生长状态；（2）MTT法测定细胞增殖，测定的数据应用SPSS16.0进行统计学分析，绘制细胞生长增殖曲线；（3）克隆形成实验计算克隆形成率；（4）向成骨细胞、脂肪细胞、骨骼肌细胞诱导分化；（5）流式细胞仪检测细胞表面抗原标志。结果：（1）细胞经传代后形态一致，呈梭形或三角形；（2）细胞生长曲线显示下颌骨BMMSCs经历潜伏期、对数生长期和平台期，统计数据表明细胞1—7d生长增殖有统计学意义（P＜0.05）；（3）集落形成实验克隆形成率37%；（4）成骨诱导后形成钙结节，茜素红染色呈阳性；成脂诱导细胞核周围出现脂滴，油红O染色呈阳性；成骨骼肌细胞诱导desmin抗原标记阳性；（5）流式细胞仪检测结果显示所获得的下颌骨BMMSCs纯度较高，CD90和CD146表面标记阳性率分别为98.7%、98.1%。结论：兔下颌骨分离出的BMMSCs，纯度较高，有强大的自我复制及增殖能力，具有多向诱导分化的潜能。
[Abstract]:Aim: to investigate the isolation, culture and biological behavior of bone marrow mesenchymal stem cells (marrow mesenchymal stemcells, BMMSCs) derived from rabbit mandible in vitro. Methods: the cancellous bone was collected from rabbit mandible and mononuclear cells were collected and cultured by density gradient centrifugation. The data were analyzed statistically by SPSS 16.0. The clone forming rate was calculated by the clone forming experiment. The colony formation rate was calculated. The clone formation rate was 4) differentiation into osteoblasts, adipocytes and skeletal muscle cells. Cell surface antigen markers were detected by flow cytometry. Results the cell growth curve showed that the mandibular BMMSCs experienced latent period, logarithmic growth period and plateau phase. The statistical data showed that the proliferation of BMMSCs in 1-7 days was statistically significant (P < 0.05). The colony forming rate of colony formation was 37%. After osteogenesis, calcium nodules were formed, alizarin red staining was positive, lipid droplets appeared around the nucleus of adipogenic cells, oil red O staining was positive, skeletal muscle cells induced desmin antigen labeling. The results of flow cytometry showed that the positive rates of CD90 and CD146 on the surface of BMMSCs were 98.7and 98.1, respectively. Conclusion: BMMSCs isolated from rabbit mandible have high purity, strong ability of self-replication and proliferation, and have the potential to induce differentiation.
【学位授予单位】：佳木斯大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R329

【参考文献】