转化生长因子-β1对脂肪干细胞增殖和Ⅰ型胶原产生的影响
发布时间：2019-02-21 13:38
【摘要】： 目的 探讨脂肪干细胞的分离培养增殖,胶原的产生和转化生长因子—β1对其增殖和胶原产生的影响。 方法 从鼠腹股沟区脂肪分离出干细胞并培养,通过倒置相差显微镜观察细胞的形态、生长增殖情况；并描绘细胞生长曲线;用流式细胞仪对细胞表面标志物进行检测。将培养的细胞分为4个组,分别加入5μg/L, 10μg/L的TGF-β1,对照组不加TGF-β1,培养后,检测细胞的数量和胶原产生量,并与使用不含TGF-β1培养的对照组及不同浓度TGF-β1组之间比较。 结果 脂肪组织中确实存在MSCs,其分离成功率为56%；通过倒置相差显微镜观察,发现脂肪来源的MSCs呈贴壁生长,外观为成纤维细胞样外观,细胞排列呈螺旋状；生长曲线显示无论原代还是传代培养的脂肪来源的MSCs的生长曲线都为S型,其中细胞增殖能力最强的是第3代和第5代；此外,AMSCs可以产生Ⅰ型胶原组织,TGF-β1在一定浓度范围内对AMSCs增殖数目的影响无显著性差异,但能显著性地增加Ⅰ型胶原组织产生(P0.05)。流式细胞仪检测结果显示,UCB-MSCs均稳定的表达与MSCs相关的表面抗原标志物CD29、CD44和CD90等,不表达造血标志CD34和CD45。 结论 MSCs可从脂肪组织中分离并培养,其成功率较高；脂肪来源的间充质干细胞的生长特性类似于呈纤维母细胞；其具有和骨髓MSCs相似的细胞表面标志物、形态学特征以及生长增殖特点等生物学特性,自我更新能力生长增殖能力强大。TGF-β1能够促进调节TGF-β1的水平能调节胶原组织的产生,可能为临床上促进肌腱愈合并防止肌腱粘连提供新的途径。
[Abstract]:Objective to investigate the proliferation of adipose stem cells, the production of collagen and the effect of transforming growth factor-尾 1 on the proliferation and collagen production of adipose stem cells. Methods Stem cells were isolated from rat inguinal fat and cultured. The morphology and proliferation of the cells were observed by inverted phase contrast microscope. Cell growth curve was described and cell surface markers were detected by flow cytometry. The cultured cells were divided into four groups, which were treated with 5 渭 g / L and 10 渭 g / L TGF- 尾 _ 1, respectively, while the control group was not treated with TGF- 尾 _ 1. After cultured, the number of cells and the amount of collagen produced were measured. The results were compared with control group without TGF- 尾 1 and TGF- 尾 1 group with different concentrations. Results MSCs, did exist in adipose tissue and the success rate of separation was 56%. By using inverted phase contrast microscope, it was found that MSCs derived from fat was adherent to the wall, and the appearance was fibroblast-like, and the cells were arranged in a spiral shape. The growth curve showed that the growth curve of MSCs from both primary and subculture fat sources was S-type, and the cell proliferation ability was the highest in the third and fifth generation. In addition, AMSCs could produce type I collagen tissue, and TGF- 尾 1 had no significant effect on the proliferation of AMSCs in a certain concentration range, but could significantly increase the production of type 鈪，
本文编号：2427555
[Abstract]:Objective to investigate the proliferation of adipose stem cells, the production of collagen and the effect of transforming growth factor-尾 1 on the proliferation and collagen production of adipose stem cells. Methods Stem cells were isolated from rat inguinal fat and cultured. The morphology and proliferation of the cells were observed by inverted phase contrast microscope. Cell growth curve was described and cell surface markers were detected by flow cytometry. The cultured cells were divided into four groups, which were treated with 5 渭 g / L and 10 渭 g / L TGF- 尾 _ 1, respectively, while the control group was not treated with TGF- 尾 _ 1. After cultured, the number of cells and the amount of collagen produced were measured. The results were compared with control group without TGF- 尾 1 and TGF- 尾 1 group with different concentrations. Results MSCs, did exist in adipose tissue and the success rate of separation was 56%. By using inverted phase contrast microscope, it was found that MSCs derived from fat was adherent to the wall, and the appearance was fibroblast-like, and the cells were arranged in a spiral shape. The growth curve showed that the growth curve of MSCs from both primary and subculture fat sources was S-type, and the cell proliferation ability was the highest in the third and fifth generation. In addition, AMSCs could produce type I collagen tissue, and TGF- 尾 1 had no significant effect on the proliferation of AMSCs in a certain concentration range, but could significantly increase the production of type 鈪，
本文编号：2427555
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