本文選題：肝細(xì)胞 + 連接蛋白　； 參考：《中國(guó)組織工程研究》2015年14期

【摘要】：背景:胎肝干細(xì)胞具有分化成肝細(xì)胞、膽管細(xì)胞的潛能,參與肝臟的修復(fù)與重建,是肝細(xì)胞的重要來(lái)源,但是人體的胎肝干細(xì)胞含量極少,如何獲取一定數(shù)量和較高純度的胎肝干細(xì)胞是目前研究的熱點(diǎn)。目的:構(gòu)建能有效抑制大鼠胎肝干細(xì)胞Cx43基因表達(dá)的siR NA載體,探討抑制Cx43表達(dá)對(duì)體外培養(yǎng)的鼠胎肝干細(xì)胞增殖及細(xì)胞周期的影響。方法:體外培養(yǎng)鼠胎肝干細(xì)胞,設(shè)計(jì)及合成靶向Cx43的siR NA序列(Cx43-siR NA)以及陰性對(duì)照序列(NC-si RNA),采用電轉(zhuǎn)法轉(zhuǎn)染大鼠胎肝干細(xì)胞,即為實(shí)驗(yàn)組和對(duì)照組,未轉(zhuǎn)染的胎肝干細(xì)胞為空白組。應(yīng)用real-time PCR和Western blot法檢測(cè)轉(zhuǎn)染前后鼠胎肝干細(xì)胞Cx43基因和蛋白的表達(dá);細(xì)胞生長(zhǎng)曲線、CCK-8法觀察細(xì)胞生長(zhǎng)增殖情況;流式細(xì)胞術(shù)測(cè)定細(xì)胞周期分布的變化。結(jié)果與結(jié)論:轉(zhuǎn)染Cx43-siR NA后,實(shí)驗(yàn)組與對(duì)照組、空白組相比Cx43基因和蛋白水平表達(dá)均明顯降低,細(xì)胞的生長(zhǎng)速度明顯增快,G0/G1期細(xì)胞減少,S期細(xì)胞數(shù)增多,差異有顯著性意義(P0.05),結(jié)果表明通過(guò)電轉(zhuǎn)法轉(zhuǎn)染靶向Cx43的si RNA能促進(jìn)體外培養(yǎng)的鼠胎肝干細(xì)胞增殖,對(duì)其培養(yǎng)有優(yōu)化作用。
[Abstract]:Background: fetal liver stem cells (FHSCs) have the potential to differentiate into hepatocytes and bile duct cells and participate in the repair and reconstruction of liver, which is an important source of hepatocytes. How to obtain a certain number and high purity of fetal liver stem cells is a hot topic at present. Aim: to construct a siR na vector that can effectively inhibit the expression of Cx43 gene in rat fetal liver stem cells (FHSCs), and to investigate the effect of inhibiting Cx43 expression on the proliferation and cell cycle of rat fetal liver stem cells (FHSCs) cultured in vitro. Methods: rat fetal liver stem cells were cultured in vitro. The siR na sequence (Cx43-siR NAA) targeting Cx43 and the negative control sequence (NC-si RNAs) were designed and synthesized. Rat fetal liver stem cells were transfected by electrotransposed method, that is, the experimental group and the control group. The untransfected fetal liver stem cells were blank group. Real-time PCR and Western blot methods were used to detect the expression of Cx43 gene and protein in fetal liver stem cells before and after transfection, cell growth curve CCK-8 method was used to observe cell growth and proliferation, and cell cycle distribution was measured by flow cytometry. Results and conclusion: after transfection of Cx43-siR na, the expression of Cx43 gene and protein in the experimental group and the control group were significantly lower than those in the control group, and the cell growth rate was significantly increased. The results showed that si RNA targeting Cx43 could promote the proliferation of rat fetal liver stem cells in vitro and optimize its culture.
【作者單位】： 天津市第四中心醫(yī)院肝膽外科;天津市南開醫(yī)院微創(chuàng)外科;
【分類號(hào)】：R575

【參考文獻(xiàn)】

相關(guān)期刊論文 前7條

1 黃利華;郭姣;黃s，

本文編號(hào)：1980091