大鼠脂肪间充质干细胞和骨髓间充质干细胞成骨分化比较的体外研究

发布时间：2018-07-31 19:08

【摘要】：目的：分离大鼠骨髓间充质干细胞和脂肪间充质干细胞，比较两种细胞的特征，为脂肪间充质干细胞可替代骨髓基质细胞做为种子细胞提供依据。方法：利用细胞贴壁性对骨髓间充质干细胞和脂肪间充质干细胞进行分离，比较两种细胞的情况，包括(1)单位质量组织获得两种细胞的量、(2)细胞形态、(3)细胞贴壁率、(4)细胞动力学、(5)细胞表面标记。并对数据进行分析。结果：(1)单位质量组织获得骨髓间充质干细胞数量为(2.23±0.35)×10~5／g，脂肪间充质干细胞为(2.66±0.75)×10~5/g，采用成组设计两样本比较的t检验，t=-1.446，P=0.179＞0.05，尚不能认为两者有差异，可以认为在单位质量骨髓和脂肪组织中获得的骨髓间充质干细胞和脂肪间充质干细胞数量相当。(2)两种细胞形态均为条索样，呈成纤维细胞形态。(3)应用直线相关分析考察两组细胞的贴壁率之间的关系，相关系数r=0.999，决定系数r~2=0.997，(F=5862.949，，P＜0.001)，可以认为两贴壁率之间有直线相关关系，可以认为在不同的时间点两种细胞的贴壁率相似，并且有共同递增趋势。(4)脂肪间充质干细胞的增殖能力与骨髓间充质干细胞相当，消化后的第2、5代细胞经历24～48小时的潜伏期后，进入6～9天的对数生长期，然后过渡到平台期。(5)通过流式实验和免疫细胞化学显示，骨髓间充质干细胞和骨髓间充质干细胞表面表达CD29、CD44，不表达CD34。结论：(1)自大鼠脂肪中可提取出与骨髓间充质干细胞类似的脂肪间充质干细胞。(2)脂肪间充质干细胞的表面标记与骨髓间充质干
[Abstract]:Aim: to isolate rat bone marrow mesenchymal stem cells (BMSCs) and adipose mesenchymal stem cells (AMSCs) and compare the characteristics of the two cells so as to provide a basis for replacing bone marrow stromal cells (BMSCs) with adipose mesenchymal stem cells (AMSCs) as seed cells. Methods: bone marrow mesenchymal stem cells (BMSCs) and adipose mesenchymal stem cells (AMSCs) were isolated with adherent cells. It includes (1) the quantity of two kinds of cells obtained by unit mass tissue, (2) cell morphology, (3) cell adhesion rate, (4) cell dynamics, and (5) cell surface labeling. The data are analyzed. Results: (1) the number of bone marrow mesenchymal stem cells (BMSCs) and adipose mesenchymal stem cells (BMSCs) were (2.23 卤0.35) 脳 10 ~ (5) / g and (2.66 卤0.75) 脳 10 ~ (5) / g respectively. It can be concluded that the number of bone marrow mesenchymal stem cells and adipose mesenchymal stem cells obtained in bone marrow and adipose tissue per unit mass is equal. (3) the linear correlation analysis was used to investigate the relationship between the adherent rate of the two groups, the correlation coefficient was 0.999, the determining coefficient was 0.997, and the coefficient of determination was 0.997 (F _ (5862.949) P < 0.001). It can be concluded that there is a linear correlation between the two cell adhesion rates. It can be concluded that the adhesion rate of the two cells at different time points is similar and there is a common increasing trend. (4) the proliferation ability of adipose mesenchymal stem cells is similar to that of bone marrow mesenchymal stem cells. After digestion, the second passage 5 cells experienced the incubation period of 24 hours 48 hours, entered the logarithmic growth phase of 6 to 9 days, and then transitioned to the plateau phase. (5) flow cytometry and immunocytochemistry showed that, Bone marrow mesenchymal stem cells and bone marrow mesenchymal stem cells expressed CD29 CD44, not CD34. Conclusion: (1) adipose mesenchymal stem cells similar to bone marrow mesenchymal stem cells can be extracted from fat of rats. (2) Surface markers of adipose mesenchymal stem cells and bone marrow mesenchymal stem cells can be obtained.
【学位授予单位】：中南大学
【学位级别】：博士
【学位授予年份】：2006
【分类号】：R329.2

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