本文選題：間質(zhì)干細(xì)胞 + 流式細(xì)胞術(shù)��； 參考：《中國組織工程研究》2017年13期

【摘要】：背景:前期已完成體外定向誘導(dǎo)骨髓間充質(zhì)干細(xì)胞分化為心肌樣細(xì)胞的實(shí)驗(yàn)。目的:觀察骨髓間充質(zhì)干細(xì)胞分化為心肌樣細(xì)胞過程中,細(xì)胞形態(tài)變化、結(jié)構(gòu)發(fā)生規(guī)律與GATA-4、Nkx-2.5和a-MHC表達(dá)的關(guān)聯(lián)性。方法:采用心肌組織裂解液誘導(dǎo)SD乳鼠骨髓間充質(zhì)干細(xì)胞分化為心肌細(xì)胞,以免疫組織化學(xué)染色檢測心肌肌鈣蛋白T和間隙連接蛋白43表達(dá)情況,鑒定心肌細(xì)胞;在定向誘導(dǎo)分化過程中,RT-PCR檢測GATA-4、Nkx2.5與α-MHC的表達(dá)情況。結(jié)果與結(jié)論:(1)細(xì)胞形態(tài):在定向誘導(dǎo)過程中,細(xì)胞形態(tài)由長梭形經(jīng)歷短桿狀、形成短小突起、網(wǎng)狀結(jié)構(gòu)、呈竹節(jié)狀及肌管樣結(jié)構(gòu);(2)心肌細(xì)胞鑒定:竹節(jié)樣結(jié)構(gòu)開始出現(xiàn)心肌肌鈣蛋白T和間隙連接蛋白43陽性細(xì)胞,肌管樣結(jié)構(gòu)心肌肌鈣蛋白T和間隙連接蛋白43陽性細(xì)胞增多;(3)RT-PCR檢測:在定向分化過程中,細(xì)胞GATA-4、Nkx2.5與α-MHC基因表達(dá)持續(xù)增加;細(xì)胞交錯(cuò)連接成網(wǎng)狀時(shí),GATA-4表達(dá)明顯增加,之后持續(xù)高表達(dá);相鄰細(xì)胞融合形成肌管樣結(jié)構(gòu)時(shí),α-MHC表達(dá)達(dá)到峰值;Nkx2.5表達(dá)隨誘導(dǎo)時(shí)間的延長逐漸增加;(4)結(jié)果表明:骨髓間充質(zhì)干細(xì)胞分化為心肌樣細(xì)胞過程中,心肌細(xì)胞特異基因的表達(dá)與心肌樣細(xì)胞的形態(tài)特征、特殊結(jié)構(gòu)發(fā)生密切相關(guān),具有典型的時(shí)序性和空間性。
[Abstract]:Background: the in vitro induction of bone marrow mesenchymal stem cells into cardiomyocyte-like cells has been completed. Aim: to investigate the relationship between the morphological changes of bone marrow mesenchymal stem cells (BMSCs) and the expression of GATA-4nkx-2.5 and a-MHC during differentiation into cardiomyocyte-like cells. Methods: bone marrow mesenchymal stem cells from neonatal SD rats were induced to differentiate into cardiomyocytes by myocardial tissue lysate. The expression of cardiac troponin T and gap junction protein 43 were detected by immunohistochemical staining to identify cardiac myocytes. The expression of GATA-4, Nkx2.5 and 偽 -MHC was detected by RT-PCR during directed differentiation. Results and conclusion: in the process of directional induction, the cell morphology was from long fusiform to short rod shape, forming short protuberance and reticular structure. Cardiac myocyte identification: cardiac troponin T and gap junction 43 positive cells began to appear in bamboo gangliform structure. The expression of GATA-4Nkx2.5 and 偽 -MHC gene in cardiac troponin T and interstitial connexin 43 positive cells was continuously increased during the directional differentiation, and the expression of GATA-4 was significantly increased when the cells were interlaced into reticulation. The expression of 偽 -MHC increased gradually with the prolongation of induction time. The results showed that during the differentiation of bone marrow mesenchymal stem cells into cardiomyocyte-like cells, the expression of 偽 -MHC reached a peak when the adjacent cells fused to form myocyte-like structure, and the expression of Nkx2.5 increased gradually with the prolongation of induction time. The expression of cardiomyocyte specific genes is closely related to the morphologic characteristics and special structure of cardiomyoid cells, and has typical temporal and spatial characteristics.
【作者單位】： 廣西醫(yī)科大學(xué)組織學(xué)與胚胎學(xué)教研室;
【基金】：廣西急診與醫(yī)學(xué)救援人才小高地·廣西高校急診醫(yī)學(xué)重點(diǎn)實(shí)驗(yàn)室開放課題(GXJZ201505) 廣西醫(yī)科大學(xué)青年科學(xué)基金資助項(xiàng)目(GXMUYSF201528) 廣西自然科學(xué)基金(2012GXNSFAA053126,桂科自0832157,桂科攻0592007-1G,桂科青0229009) 廣西高校人體發(fā)育與疾病研究重點(diǎn)實(shí)驗(yàn)(0102402214009C)~~
【分類號】：R329.2

【相似文獻(xiàn)】

相關(guān)期刊論文 前10條

1 法憲恩;王利霞;侯劍峰;張瑞成;王海永;;5-氮胞苷誘導(dǎo)骨髓間充質(zhì)干細(xì)胞向心肌樣細(xì)胞分化觀察[J];鄭州大學(xué)學(xué)報(bào)(醫(yī)學(xué)版);2005年06期

2 胡波,張中明,董紅燕,張好生;大鼠骨髓基質(zhì)細(xì)胞體外培養(yǎng)誘導(dǎo)為心肌樣細(xì)胞的實(shí)驗(yàn)研究[J];徐州醫(yī)學(xué)院學(xué)報(bào);2002年02期

3 銀廣悅;張繼領(lǐng);陳素萍;張龍;張明;;人骨髓間充質(zhì)干細(xì)胞體外誘導(dǎo)分化為心肌樣細(xì)胞及鑒定[J];中華實(shí)用診斷與治療雜志;2012年08期

4 方志成;王瑋;黃從新;王為民;何朝榮;;骨髓間(充)質(zhì)干細(xì)胞體外定向誘導(dǎo)心肌樣細(xì)胞超微結(jié)構(gòu)特征研究[J];心血管康復(fù)醫(yī)學(xué)雜志;2005年06期

5 程芮;王士雯;曹豐;孟玲;王韞芳;李艷華;李泱;裴雪濤;;5-氮胞苷濃度對體外骨髓間充質(zhì)干細(xì)胞誘導(dǎo)分化為心肌樣細(xì)胞的影響[J];中華老年多器官疾病雜志;2005年03期

6 王紅祥;李賓公;邵詩穎;唐曉瓊;趙智剛;鄒萍;;脂肪基質(zhì)干細(xì)胞誘導(dǎo)分化為心肌樣細(xì)胞的實(shí)驗(yàn)研究[J];華中科技大學(xué)學(xué)報(bào)(醫(yī)學(xué)版);2006年05期

7 馬新U，

本文編號：1936552