發(fā)布時間：2019-02-21 13:38

【摘要】： 目的 探討脂肪干細(xì)胞的分離培養(yǎng)增殖,膠原的產(chǎn)生和轉(zhuǎn)化生長因子—β1對其增殖和膠原產(chǎn)生的影響。 方法 從鼠腹股溝區(qū)脂肪分離出干細(xì)胞并培養(yǎng),通過倒置相差顯微鏡觀察細(xì)胞的形態(tài)、生長增殖情況；并描繪細(xì)胞生長曲線;用流式細(xì)胞儀對細(xì)胞表面標(biāo)志物進(jìn)行檢測。將培養(yǎng)的細(xì)胞分為4個組,分別加入5μg/L, 10μg/L的TGF-β1,對照組不加TGF-β1,培養(yǎng)后,檢測細(xì)胞的數(shù)量和膠原產(chǎn)生量,并與使用不含TGF-β1培養(yǎng)的對照組及不同濃度TGF-β1組之間比較。 結(jié)果 脂肪組織中確實(shí)存在MSCs,其分離成功率為56%；通過倒置相差顯微鏡觀察,發(fā)現(xiàn)脂肪來源的MSCs呈貼壁生長,外觀為成纖維細(xì)胞樣外觀,細(xì)胞排列呈螺旋狀；生長曲線顯示無論原代還是傳代培養(yǎng)的脂肪來源的MSCs的生長曲線都為S型,其中細(xì)胞增殖能力最強(qiáng)的是第3代和第5代；此外,AMSCs可以產(chǎn)生Ⅰ型膠原組織,TGF-β1在一定濃度范圍內(nèi)對AMSCs增殖數(shù)目的影響無顯著性差異,但能顯著性地增加Ⅰ型膠原組織產(chǎn)生(P0.05)。流式細(xì)胞儀檢測結(jié)果顯示,UCB-MSCs均穩(wěn)定的表達(dá)與MSCs相關(guān)的表面抗原標(biāo)志物CD29、CD44和CD90等,不表達(dá)造血標(biāo)志CD34和CD45。 結(jié)論 MSCs可從脂肪組織中分離并培養(yǎng),其成功率較高；脂肪來源的間充質(zhì)干細(xì)胞的生長特性類似于呈纖維母細(xì)胞；其具有和骨髓MSCs相似的細(xì)胞表面標(biāo)志物、形態(tài)學(xué)特征以及生長增殖特點(diǎn)等生物學(xué)特性,自我更新能力生長增殖能力強(qiáng)大。TGF-β1能夠促進(jìn)調(diào)節(jié)TGF-β1的水平能調(diào)節(jié)膠原組織的產(chǎn)生,可能為臨床上促進(jìn)肌腱愈合并防止肌腱粘連提供新的途徑。
[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，

本文編號：2427555