本文選題：接觸式共培養(yǎng) + 間充質(zhì)干細(xì)胞��； 參考：《中國(guó)脊柱脊髓雜志》2014年06期

【摘要】：目的:比較兩種人間充質(zhì)干細(xì)胞在接觸式共培養(yǎng)體系下對(duì)退變髓核細(xì)胞生物學(xué)功能的影響。方法:取同一例腰椎間盤突出癥患者的退變髓核組織、脂肪組織和骨髓組織,分別分離培養(yǎng)退變髓核細(xì)胞(NPCs)、脂肪間充質(zhì)干細(xì)胞(ADSCs)和骨髓間充質(zhì)干細(xì)胞(BMSCs)。取3種細(xì)胞傳代至第3代的細(xì)胞進(jìn)行接觸式共培養(yǎng)。共培養(yǎng)前,利用PKH26對(duì)第3代退變NPCs進(jìn)行染色標(biāo)記,然后按1∶1的細(xì)胞比例,將NPCs分別與ADSCs和BMSCs在普通6孔板內(nèi)進(jìn)行接觸式共培養(yǎng)。實(shí)驗(yàn)分為3組,A組為單獨(dú)培養(yǎng)的NPCs,為對(duì)照組;B組為與ADSCs共培養(yǎng)的NPCs;C組為與BMSCs共培養(yǎng)的NPCs。共培養(yǎng)第7天時(shí),利用MoFlo高速流式細(xì)胞分選儀將共培養(yǎng)的細(xì)胞進(jìn)行分選,獲取各組NPCs。然后提取各組NPCs的總RNA,進(jìn)行反轉(zhuǎn)錄后,再利用Real-Time PCR技術(shù)檢測(cè)各組NPCs的Ⅱ型膠原、蛋白多糖和SOX-9基因的相對(duì)表達(dá)量。將Ⅱ型膠原、蛋白多糖和SOX-9基因的相對(duì)表達(dá)量按分組進(jìn)行組間t檢驗(yàn),P0.05為有統(tǒng)計(jì)學(xué)差異。結(jié)果:從退變髓核組織、骨髓組織和脂肪組織中,均成功分離培養(yǎng)出NPCs、BMSCs和ADSCs。待各細(xì)胞傳至第3代時(shí),成功利用PKH26對(duì)NPCs進(jìn)行染色標(biāo)記。按實(shí)驗(yàn)分組將各細(xì)胞進(jìn)行接觸式共培養(yǎng)后第7天,成功利用MoFlo高速流式細(xì)胞分選儀將共培養(yǎng)細(xì)胞進(jìn)行分選,獲取各組NPCs。經(jīng)提取各組NPCs的總RNA,反轉(zhuǎn)錄并進(jìn)行Real-time PCR后,獲取各組NPCs的Ⅱ型膠原、蛋白多糖和SOX-9基因相對(duì)表達(dá)量,A組分別為1.03±0.04,1.05±0.07,1.04±0.17;B組分別為5.26±0.24,7.71±0.21,3.84±0.25;C組分別為9.33±0.39,11.07±0.34,5.64±0.26;B、C組NPCs的Ⅱ型膠原、蛋白多糖和SOX-9基因相對(duì)表達(dá)量與A組比較顯著性增加(P0.05);C組與B組比較,各指標(biāo)基因相對(duì)表達(dá)量顯著性增加(P0.05)。結(jié)論:在接觸式共培養(yǎng)條件下,BMSCs和ADSCs對(duì)退變NPCs均具有營(yíng)養(yǎng)激活效應(yīng);BMSCs對(duì)退變NPCs的激活效應(yīng)比ADSCs更好,對(duì)于椎間盤退行性疾病的生物學(xué)治療可能更具優(yōu)勢(shì)。
[Abstract]:Aim: to compare the effects of two human mesenchymal stem cells on the biological function of degenerative nucleus pulposus cells in contact coculture system. Methods: the degenerative nucleus pulposus tissue, adipose tissue and bone marrow tissue were isolated from the same patient with lumbar disc herniation. NPCsm cells, adipose mesenchymal stem cells (ADSCss) and bone marrow mesenchymal stem cells (BMSCs) were isolated and cultured respectively. Three kinds of cells were subcultured to the third passage for contact coculture. Before co-culture, the third generation degenerative NPCs was stained with PKH26, and then NPCs and ADSCs and BMSCs were co-cultured in ordinary 6-well plate according to the ratio of 1:1 cells. The experiment was divided into 3 groups: group A was isolated NPCs and group B was co-cultured with ADSCs. Group C was co-cultured with BMSCs. On the 7th day of co-culture, the co-cultured cells were separated by MoFlo high speed flow cytometry to obtain each group of NPCs. Then the total RNAs of each group were extracted, and then the relative expression of type 鈪，

本文編號(hào)：1894081