本文選題：骨髓　切入點(diǎn)：間質(zhì)干細(xì)胞　出處：《中國(guó)組織工程研究》2017年25期 　論文類(lèi)型：期刊論文

【摘要】：背景:骨髓間充質(zhì)干細(xì)胞來(lái)源有限,而且在多次體外傳代培養(yǎng)過(guò)程中,其細(xì)胞形態(tài)、增殖及多向分化能力會(huì)發(fā)生改變。目的:探討成纖維細(xì)胞生長(zhǎng)因子對(duì)經(jīng)過(guò)多次傳代、細(xì)胞形態(tài)發(fā)生改變的骨髓間充質(zhì)干細(xì)胞形態(tài)、增殖及多向分化能力的影響。方法:(1)體外分離培養(yǎng)大鼠骨髓間充質(zhì)干細(xì)胞,連續(xù)傳代培養(yǎng)6次,觀察其細(xì)胞形態(tài)變化;(2)將第6代細(xì)胞接種于96孔板中,隨機(jī)分為對(duì)照組和成纖維細(xì)胞生長(zhǎng)因子處理組,相應(yīng)培養(yǎng)1,2,3,4,5,6,7 d后,用CCK-8試劑盒檢測(cè)細(xì)胞增殖狀況;(3)將第6代細(xì)胞接種于6孔板中,隨機(jī)分為對(duì)照組和成纖維細(xì)胞生長(zhǎng)因子處理組,分別用普通培養(yǎng)基和含成纖維細(xì)胞生長(zhǎng)因子培養(yǎng)基培養(yǎng)7 d,換成骨、成脂、成軟骨誘導(dǎo)培養(yǎng)基繼續(xù)培養(yǎng)7 d,應(yīng)用熒光定量PCR法檢測(cè)成骨(RUNX2、ALP、OCN)、成脂(PPARγ2、AP2、ADIPOQ)、成軟骨(SOX9、Collagen Ⅱ、aggrecan)相關(guān)基因的表達(dá),應(yīng)用蛋白印記法檢測(cè)RUNX2、PPARγ2、SOX9蛋白的表達(dá);(4)將第6代細(xì)胞接種到6孔細(xì)胞培養(yǎng)板中,隨機(jī)分為對(duì)照組及成纖維細(xì)胞生長(zhǎng)因子處理組,分別用普通生長(zhǎng)培養(yǎng)基及含成纖維細(xì)胞生長(zhǎng)因子的生長(zhǎng)培養(yǎng)基培養(yǎng)7 d,換用成骨、成脂、成軟骨誘導(dǎo)培養(yǎng)基繼續(xù)培養(yǎng)14 d,進(jìn)行茜素紅染色、油紅O染色和阿利新藍(lán)染色。結(jié)果與結(jié)論:(1)經(jīng)過(guò)連續(xù)6次傳代培養(yǎng),骨髓間充質(zhì)干細(xì)胞形態(tài)發(fā)生明顯變化,成纖維細(xì)胞生長(zhǎng)因子處理7 d后其形態(tài)逐漸恢復(fù)原代特性;(2)與對(duì)照組相比,培養(yǎng)第3-7天成纖維細(xì)胞生長(zhǎng)因子處理組的細(xì)胞增殖速度明顯加快,差異有顯著性意義(P0.05);(3)與對(duì)照組相比,成纖維細(xì)胞生長(zhǎng)因子處理組細(xì)胞成骨相關(guān)基因(RUNX2,ALP,OCN)、成脂相關(guān)基因(PPARγ2,AP2,ADIPOQ)、成軟骨相關(guān)基因(SOX9,collagen 2,aggrecan)表達(dá)明顯升高,差異有顯著性意義(P0.05);(4)成纖維細(xì)胞生長(zhǎng)因子處理組RUNX2、PPARγ2、SOX9蛋白表達(dá)明顯高于對(duì)照組(P0.05);(5)與對(duì)照組相比,成纖維細(xì)胞生長(zhǎng)因子預(yù)處理組細(xì)胞產(chǎn)生胞外鈣結(jié)節(jié)的數(shù)量、細(xì)胞內(nèi)脂滴的數(shù)量、細(xì)胞酸性黏多糖的表達(dá)明顯增加;(6)結(jié)果表明,成纖維細(xì)胞生長(zhǎng)因子可以維持多次傳代骨髓間充質(zhì)干細(xì)胞的干細(xì)胞特性。
[Abstract]:Background: bone marrow mesenchymal stem cells (BMSCs) have limited sources, and their cell morphology, proliferation and multidirectional differentiation ability will change in vitro. Objective: to investigate the effects of fibroblast growth factor (FGF) on the proliferation and differentiation of bone marrow mesenchymal stem cells (BMSCs). Effects of Morphogenetic changes on the Morphology, Proliferation and Multidirectional differentiation of Bone Marrow Mesenchymal Stem cells methods Rat Bone Marrow Mesenchymal Stem cells were isolated and cultured in vitro for 6 consecutive times. The sixth passage cells were inoculated into 96 well plate and randomly divided into control group and fibroblast growth factor treated group. The sixth passage cells were inoculated into 6-well plate with CCK-8 kit. The cells were randomly divided into two groups: control group and fibroblast growth factor treatment group. Normal culture medium and fibroblast growth factor medium were used for 7 days, then bone and fat were replaced, and cartilage induction medium was cultured for 7 days. Fluorescence quantitative PCR was used to detect the expression of genes related to osteoblasts RUNX2, APPAR- 緯 2, AP2, ADIPOQN and cartilage SOX9Collagen 鈪，

本文編號(hào)：1557594