發(fā)布時(shí)間：2018-05-20 02:29

  本文選題：臍帶 + 間充質(zhì)干細(xì)胞��； 參考：《汕頭大學(xué)》2007年碩士論文

【摘要】： 目的：探討人臍帶間充質(zhì)干細(xì)胞(MSCs)向心肌細(xì)胞分化的可能性，為心肌細(xì)胞的移植探索新的細(xì)胞來(lái)源。 方法：從人臍帶華爾通氏膠分離、培養(yǎng)MSCs，，用(5-azacytidine，5-aza)和二甲基亞砜(DMSO)誘導(dǎo)人臍帶間充質(zhì)干細(xì)胞的分化。觀察誘導(dǎo)后分化細(xì)胞形態(tài)的變化，用免疫細(xì)胞化學(xué)方法檢測(cè)分化的細(xì)胞是否表達(dá)心肌肌鈣蛋白I(troponin I)、心肌肌鈣蛋白T(troponinT)和心肌結(jié)蛋白(desmin)，RT-PCR方法測(cè)定分化的細(xì)胞是否有心肌特異轉(zhuǎn)錄因子(Nkx2.5)和心肌細(xì)胞結(jié)蛋白(desmin)的cDNA的表達(dá)。透射電鏡觀察分化的心肌樣細(xì)胞內(nèi)是否有肌絲的存在，掃描電鏡觀察分化的心肌樣細(xì)胞的形態(tài)變化，從不同的層面鑒定誘導(dǎo)分化的心肌樣細(xì)胞是否具有心肌細(xì)胞的特性和功能? 結(jié)果：從人臍帶華爾通膠分離、培養(yǎng)的貼壁細(xì)胞，體外生長(zhǎng)形態(tài)類似于成纖維細(xì)胞，可以維持在未分化狀態(tài)穩(wěn)定增殖，體外增殖超過(guò)10代，細(xì)胞凍存一個(gè)月后復(fù)蘇，生長(zhǎng)特點(diǎn)與凍存前基本一致。人臍帶MSCs經(jīng)5-aza和DMSO誘導(dǎo)后可向心肌樣細(xì)胞分化，分化的細(xì)胞表達(dá)心肌細(xì)胞的標(biāo)記troponin I、troponinT和desmin。RT-PCR檢測(cè)證實(shí)，人臍帶MSCs誘導(dǎo)前不表達(dá)Nkx2.5和desmin，經(jīng)5-aza和DMSO誘導(dǎo)后，分化的細(xì)胞檢測(cè)到有心肌特異轉(zhuǎn)錄因子(Nkx2.5)和心肌細(xì)胞結(jié)蛋白(desmin)的cDNA的表達(dá)。透射電鏡可以觀察分化的細(xì)胞漿中有肌絲的存在。 結(jié)論：人臍帶間充質(zhì)干細(xì)胞可以分化為心肌樣細(xì)胞，5-aza和DMSO可以作為心肌細(xì)胞誘導(dǎo)劑，人臍帶間充質(zhì)干細(xì)胞可以作為心肌細(xì)胞的來(lái)源。
[Abstract]:Aim: to explore the possibility of differentiation of human umbilical cord mesenchymal stem cells (MSCs) into cardiomyocytes and explore a new source of cells for cardiac myocyte transplantation. Methods: human umbilical cord mesenchymal stem cells (MSCs) were isolated from human umbilical cord and cultured with 5-azacytidine (5-aza) and dimethyl sulfoxide (DMSO) to induce the differentiation of human umbilical cord mesenchymal stem cells. The morphological changes of differentiated cells were observed after induction. Immunocytochemistry was used to detect whether differentiated cells expressed cardiac troponin I(troponin, cardiac troponin T) and cardiac desmin. RT-PCR method was used to detect whether differentiated cells had myocardial specific transcription factor Nkx2.5) and cardiomyocyte egg formation. The expression of cDNA. Transmission electron microscopy (TEM) was used to observe the presence of myofilament in differentiated cardiomyoid cells. SEM was used to observe the morphological changes of differentiated cardiomyoid cells and to identify whether the differentiated cardiomyocyte-like cells had the characteristics and functions of cardiomyocytes from different levels. Results: the adherent cells were isolated from human umbilical cord and cultured in vitro. The growth morphology of adherent cells was similar to that of fibroblasts. The cells could proliferate steadily in undifferentiated state and proliferate in vitro for more than 10 generations. The cells were resuscitated after one month of cryopreservation. The growth characteristics were basically consistent with those before freezing. Human umbilical cord MSCs could differentiate into cardiomyocyte-like cells induced by 5-aza and DMSO. The detection of troponin Itropon T and desmin.RT-PCR showed that human cord MSCs did not express Nkx2.5 and desmin before induction, but 5-aza and DMSO induced them. The expression of cDNA was detected in differentiated cells with myocardial specific transcription factor Nkx2.5) and cardiac myocyte desmin. The presence of myofilms in the differentiated cytoplasm can be observed by transmission electron microscopy (TEM). Conclusion: human umbilical cord mesenchymal stem cells can differentiate into cardiomyocyte-like cells, 5-aza and DMSO can be used as inducers of cardiomyocytes, and human umbilical cord mesenchymal stem cells can be used as sources of cardiomyocytes.
【學(xué)位授予單位】：汕頭大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2007
【分類號(hào)】：R329

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本文編號(hào)：1912809