本文選題：大鼠 + 骨髓間充質(zhì)干細(xì)胞　； 參考：《昆明醫(yī)學(xué)院》2011年碩士論文

【摘要】：[目的]：大鼠骨髓間充質(zhì)干細(xì)胞(rabbit mesenchymal stem cells, MSC)分離純化培養(yǎng)后,誘導(dǎo)為成骨細(xì)胞、心肌細(xì)胞和胰島樣細(xì)胞,觀察不同分化階段的細(xì)胞形態(tài)、檢測特殊因子或標(biāo)志物的表達變化。 [方法]：體外分離培養(yǎng)鼠骨髓間充質(zhì)干細(xì)胞,采用文獻中的誘導(dǎo)方法誘導(dǎo),分別于第3、7、14、21天倒置熒光顯微鏡動態(tài)觀察細(xì)胞形態(tài)的變化；并通過RT-PCR技術(shù)和酶聯(lián)免疫吸附技術(shù)檢測干細(xì)胞在誘導(dǎo)不同階段移植特殊因子或標(biāo)志物的表達變化。 [結(jié)果]：MSC能夠正常貼壁成長,抗原表達為CD34-/CD45-/CD73+/CD105+。倒置螢光顯微鏡觀察下細(xì)胞形態(tài)發(fā)生明顯變化,RT-PCR檢測結(jié)果顯示GATA-4、Nkx2.5、Sarcomericactin和Connexin-43mRNA向誘導(dǎo)成心肌細(xì)胞的過程中表達持續(xù)增加,Isulin 1、Isulin 2、CK-19、nestin、Glut2和pdxl mRNA表達在向胰島樣細(xì)胞誘導(dǎo)的過程中持續(xù)增加,說明在誘導(dǎo)劑誘導(dǎo)下能夠向誘導(dǎo)為成骨細(xì)胞、心肌細(xì)胞和胰島樣細(xì)胞分化。 [結(jié)論]：①建立了GFP標(biāo)記的RMSC的體外分離、培養(yǎng)及鑒定的技術(shù)平臺。②建立了MSC向心肌細(xì)胞、成骨細(xì)胞和胰島樣細(xì)胞分化的誘導(dǎo)體系。⑧RT-PCR結(jié)果顯示MSC成功向心肌細(xì)胞和胰島樣細(xì)胞分化,并分泌相應(yīng)細(xì)胞因子。④酶聯(lián)免疫吸附實驗結(jié)果顯示MSC向胰島樣細(xì)胞和成骨細(xì)胞分化,并且分泌胰島素和堿性磷酸酶。
[Abstract]:[objective]: rat bone marrow mesenchymal stem cells (BMSCs) were isolated and cultured in rabbit mesenchymal stem cells, MSC), then induced into osteoblasts, cardiomyocytes and islet like cells. [methods] Rat bone marrow mesenchymal stem cells were isolated and cultured in vitro, induced by the induction method in literature, and the morphological changes of the cells were observed by inverted fluorescence microscope on day 3, 7, 1421, respectively. RT-PCR and enzyme linked immunosorbent assay (Elisa) were used to detect the expression of specific factors or markers in different stages of stem cell transplantation. [results] CD34-/CD45-/CD73 / CD105 was expressed as CD34-/CD45-/CD73 / CD105 antigen. The results of reverse fluorescent microscopy showed that the expression of Glut2 and pdxl mRNA in islet like cells increased continuously during the process of inducing cardiac myocytes by GATA-4Nkx2.5Nkx2.5A, and the expression of Glut2 and pdxl mRNA increased continuously during the induction of cardiac myocytes, and the expression of Glut2 and pdxl mRNA increased during the induction of Glut2 and pdxl mRNA to islet like cells, and the expression of Glut2 and Glut2 in the islet like cells was increased during the induction of Glut2 and pdxl mRNA to islet like cells. These results indicated that induced osteoblasts, cardiomyocytes and islet like cells could be induced. [conclusion] the in vitro isolation, culture and identification of RMSC labeled by GFP were established in vitro. 2. 2. MSC to cardiomyocytes was established by 1: 1. The differentiation of osteoblasts and islet like cells was induced by .8RT-PCR. The results showed that MSC successfully differentiated into cardiomyocytes and islet like cells. The results of enzyme linked immunosorbent assay (Elisa) showed that MSC differentiated into islet like cells and osteoblasts and secreted insulin and alkaline phosphatase.
【學(xué)位授予單位】：昆明醫(yī)學(xué)院
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2011
【分類號】：R329

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本文編號：1908122