本文選題：肌源性干細(xì)胞　切入點(diǎn)：轉(zhuǎn)化生長因子β　出處：《中國婦產(chǎn)科臨床雜志》2015年05期

【摘要】：目的通過改良差速貼壁法分離純化綿羊MDSC,探索轉(zhuǎn)化生長因子(TGF-β1)誘導(dǎo)綿羊肌源性干細(xì)胞分化為平滑肌的可能性。方法取成年綿羊股四頭肌細(xì)胞采用改良差速貼壁法(modified preplate technique)進(jìn)行優(yōu)化、分離、純化獲得綿羊級(jí)源性干細(xì)胞(MDSC)。利用流式細(xì)胞儀鑒定CD44、CD34、CD31、CD45、CD14的表達(dá);western bloting法檢測干細(xì)胞標(biāo)志物desmin表達(dá)情況鑒定MDSC。然后利用10 ng/ml TGF-β1誘導(dǎo)MDSC向平滑肌方向分化;隨后利用Western bloting法檢測誘導(dǎo)培養(yǎng)基處理的MDSC中平滑肌蛋白標(biāo)志物a-SMA和calponin的表達(dá);結(jié)果通過對(duì)改良差速貼壁法成功分離出成年綿羊的肌源性干細(xì)胞(MDSC)。流式細(xì)胞儀鑒定其干細(xì)胞標(biāo)志物CD44、CD34陽性表達(dá),同時(shí)CD31、CD45、CD14表達(dá)陰性。Western blotting檢測MDSC中Desmin呈陽性表達(dá)。利用10 ng/ml TGF-β1成功誘導(dǎo)MDSC向平滑肌方向分化。誘導(dǎo)過程中,平滑肌特異蛋白a-SMA和calponin在誘導(dǎo)細(xì)胞中的表達(dá)逐步上升。結(jié)論本研究通過對(duì)差速貼壁法成功分離出成年雌性綿羊的肌源性干細(xì)胞(MDSC)。并首次利用TGF-β1誘導(dǎo)羊肌源性干細(xì)胞(MDSC)成功向平滑肌方向分化。
[Abstract]:Objective to study the possibility of inducing sheep myogenic stem cells to differentiate into smooth muscle cells by using modified differential adhesion method to isolate and purify sheep MDSC.Methods Adult sheep quadriceps femoris muscle cells were isolated and purified by modified differential adhesion method and induced by transforming growth factor TGF- 尾 1 (TGF- 尾 1). Modified preplate technique, Sheep grade derived stem cells were isolated and purified. The expression of CD44, CD41, CD41, CD45, CD14 was identified by flow cytometry. The expression of stem cell marker desmin was detected by western bloting. Then 10 ng/ml TGF- 尾 1 was used to induce the differentiation of MDSC into smooth muscle. The expression of smooth muscle protein markers a-SMA and calponin in MDSC treated with induction medium was detected by Western bloting assay. Results the myogenic stem cells of adult sheep were successfully isolated by modified differential adherence method. The positive expression of CD44-mCD34 was identified by flow cytometry. The expression of Desmin in MDSC was detected by Western blotting. 10 ng/ml TGF- 尾 1 was used to induce the differentiation of MDSC into smooth muscle. The expression of smooth muscle specific proteins a-SMA and calponin increased gradually. Conclusion in this study, the myogenic stem cells of adult female sheep were isolated successfully by differential adherence method. TGF- 尾 1 was used for the first time to induce the fine myogenic stem cells of sheep. MDSCs successfully differentiated into smooth muscle.
【作者單位】： 廣州醫(yī)科大學(xué)附屬第二醫(yī)院婦科;北京協(xié)和醫(yī)院婦科;
【分類號(hào)】：R711.2

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本文編號(hào)：1672631