本文選題：人骨髓間充質(zhì)干細(xì)胞 + 胰島移植��； 參考：《吉林大學(xué)》2007年博士論文

【摘要】： 為解決目前胰島移植根治糖尿病所面臨的胰島來源匿乏和免疫排斥兩大難題,本實(shí)驗(yàn)以人骨髓間充質(zhì)干細(xì)胞(hMSCs)作為胰島移植種子細(xì)胞,以適宜條件誘導(dǎo)hMSCs分化為胰島素分泌細(xì)胞并初步探討其分化機(jī)制。 以密度梯度離心法和貼壁篩選法大量培養(yǎng)擴(kuò)增hMSCs,利用高糖、尼克酰胺和Exendix-4體外誘導(dǎo)hMSCs向胰島素分泌細(xì)胞分化,通過形態(tài)學(xué)、酶學(xué)、分子生物學(xué)和電生理學(xué)等方法對誘導(dǎo)后的細(xì)胞進(jìn)行分泌功能和活性鑒定,并以PDX-1和離子通道變化為切入點(diǎn)探討其分化條件和機(jī)制。并利用BrdU標(biāo)記誘導(dǎo)后細(xì)胞移植入糖尿病裸鼠體內(nèi),進(jìn)一步觀察其是否具有分泌胰島素和降低血糖的功能。結(jié)果發(fā)現(xiàn):成功地分離培養(yǎng)并擴(kuò)增hMSCs;體外誘導(dǎo)hMSCs分化為為胰島素分泌細(xì)胞,并且分化后細(xì)胞保持良好的增殖活性,其胰島素的分泌受外環(huán)境糖濃度的調(diào)節(jié);hMSCs分化機(jī)制可能為高糖刺激了hMSCs細(xì)胞PDX-1的表達(dá)和活化以及Ca2+通道開放,從而激活下游一系列胰島分化相關(guān)基因;BrdU標(biāo)記誘導(dǎo)后細(xì)胞移植入糖尿病裸鼠體內(nèi)能夠存活增殖并具有較好的降低血糖功能,具備了一定的胰島細(xì)胞功能。 綜上所述,本實(shí)驗(yàn)系統(tǒng)全面的探討了適宜條件下hMSCs分化為胰島素分泌細(xì)胞的可能和機(jī)制,為今后更好地利用hMSCs作為胰島移植的種子細(xì)胞治療胰島素依賴型糖尿病提供了理論基礎(chǔ)和實(shí)驗(yàn)依據(jù)。
[Abstract]:In order to solve the two major problems of islet origin obscurity and immune rejection faced by islet transplantation to cure diabetes mellitus, human bone marrow mesenchymal stem cells (hMSCs) were used as seed cells for islet transplantation. HMSCs was induced to differentiate into insulin-secreting cells under suitable conditions and its differentiation mechanism was discussed. HMSCs were cultured and amplified by density gradient centrifugation and adherent screening. High glucose, nicotinamide and Exendix-4 were used to induce the differentiation of hMSCs into insulin-secreting cells in vitro. The secretory function and activity of induced cells were identified by molecular biology and electrophysiology, and the differentiation conditions and mechanism of induced cells were discussed with the changes of PDX-1 and ion channels as the starting point. The cells induced by BrdU were transplanted into nude mice to observe whether they could secrete insulin and decrease blood glucose. The results showed that hMSCs were successfully isolated and amplified, hMSCs was induced to differentiate into insulin-secreting cells in vitro, and the cells maintained good proliferative activity after differentiation. Its insulin secretion was regulated by the concentration of glucose in external environment. The mechanism of differentiation of hMSCs might be that high glucose stimulated the expression and activation of PDX-1 and the opening of Ca2 channel in hMSCs cells. As a result of activation of a series of downstream islet differentiation related genes BrdU labeled induced cells transplanted into diabetic nude mice can survive and proliferate and have a better function of lowering blood glucose and have a certain function of islet cells. In conclusion, the mechanism of hMSCs differentiation into insulin-secreting cells under suitable conditions was discussed. It provides a theoretical and experimental basis for the better use of hMSCs as seed cells for islet transplantation in the treatment of insulin-dependent diabetes.
【學(xué)位授予單位】：吉林大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2007
【分類號】：R329

【相似文獻(xiàn)】

相關(guān)期刊論文 前10條

1 李新永;;紅景天苷對人骨髓間充質(zhì)干細(xì)胞增殖及分泌的影響[J];臨床合理用藥雜志;2011年27期

2 劉國強(qiáng);張琳;洪天配;;第67屆美國糖尿病學(xué)會(huì)年會(huì)關(guān)于胰島細(xì)胞生物學(xué)和胰島移植研究進(jìn)展的綜合報(bào)道[J];中國糖尿病雜志;2007年11期

3 劉釗;邊云飛;肖傳實(shí);;晚期糖基化終產(chǎn)物通過氧化應(yīng)激引起人骨髓間充質(zhì)干細(xì)胞凋亡[J];中國動(dòng)脈硬化雜志;2011年05期

4 李偉中;邱曉燕;邱學(xué)榮;王鴻武;林麗敏;蔣學(xué)武;唐秋靈;林曉波;馬廉;;人臍帶間充質(zhì)干細(xì)胞向胰島素分泌細(xì)胞分化過程免疫特性的研究[J];中國小兒血液與腫瘤雜志;2011年04期

5 廖玉婷;李富榮;;α1-抗胰蛋白酶在胰島移植和Ⅰ型糖尿病中的作用[J];細(xì)胞與分子免疫學(xué)雜志;2011年08期

6 盧曉芳;杜晶春;李磊;賴文玉;張秀明;李偉強(qiáng);楊霞;項(xiàng)鵬;;新型慢病毒載體的構(gòu)建及其在人骨髓間充質(zhì)干細(xì)胞基因轉(zhuǎn)導(dǎo)中的應(yīng)用[J];中國組織工程研究與臨床康復(fù);2011年23期

7 王承云;石磊;夏春;鄭欣鵬;張兵;林飛太;王少杰;;人骨髓間充質(zhì)干細(xì)胞分離培養(yǎng)方法的改進(jìn)[J];中國組織工程研究與臨床康復(fù);2011年32期

8 葛煥琦;蔡寒青;門秀麗;許世清;潘煥峰;婁晉寧;;人胚胎胰腺組織干細(xì)胞的分離鑒定和純化[J];中國糖尿病雜志;2007年07期

9 祝方;鄭晟e，

本文編號：1831223