【摘要】： 間充質(zhì)干細(xì)胞(MScs)是來(lái)源于發(fā)育早期中胚層的一類多能干細(xì)胞,在誘導(dǎo)器官移植免疫耐受中有著廣闊的應(yīng)用前景。目前MSCs主要來(lái)源于成人骨髓,但許多研究顯示人臍帶是較理想的MSCs來(lái)源。關(guān)于大鼠骨髓來(lái)源的MSCs用于移植免疫耐受及進(jìn)行組織修復(fù)的研究很多,但由于沒(méi)有大鼠臍帶來(lái)源MSCs的相關(guān)研究,導(dǎo)致臍帶來(lái)源的MSCs無(wú)法進(jìn)行動(dòng)物體內(nèi)試驗(yàn)。本研究從大鼠臍帶中分離出間充質(zhì)干細(xì)胞,并對(duì)其生物學(xué)特性進(jìn)行研究,找到合適的分離培養(yǎng)方法,并進(jìn)行體外和體內(nèi)試驗(yàn),探討臍帶源間充質(zhì)干細(xì)胞減少免疫排斥反應(yīng),誘導(dǎo)免疫耐受的作用。 本研究第一部分采用膠原酶消化法和組織塊培養(yǎng)法,進(jìn)行貼壁傳代,得到大鼠臍帶源間充質(zhì)干細(xì)胞,進(jìn)行細(xì)胞表型分析和誘導(dǎo)分化。結(jié)果顯示：兩種方法得到的貼壁細(xì)胞均為間充質(zhì)干細(xì)胞,其生物學(xué)性狀與文獻(xiàn)報(bào)導(dǎo)的骨髓間充質(zhì)干細(xì)胞很接近。膠原酶消化法比組織塊培養(yǎng)法的效率更高。第二部分以Lewis大鼠脾細(xì)胞為反應(yīng)細(xì)胞,DA大鼠脾細(xì)胞為刺激細(xì)胞,加入來(lái)源于Wistar大鼠臍帶的間充質(zhì)干細(xì)胞,采用3H-胸腺嘧啶核苷摻入法,進(jìn)行混合淋巴細(xì)胞培養(yǎng),檢測(cè)大鼠骨臍帶來(lái)源充質(zhì)干細(xì)胞對(duì)體外混合淋巴細(xì)胞培養(yǎng)的影響。結(jié)果顯示臍帶來(lái)源間充質(zhì)干細(xì)胞不引起大鼠淋巴細(xì)胞增殖反應(yīng),但對(duì)大鼠混合淋巴細(xì)胞培養(yǎng)有顯著的抑制作用；第三部分以DA大鼠為供體,Lewis大鼠為受體,建立大鼠異位心臟移植模型,并于術(shù)后即刻經(jīng)尾靜脈注入Wistar大鼠臍帶來(lái)源的間充質(zhì)干細(xì)胞,探討其對(duì)大鼠同種異體心臟移植的影響。異位心臟移植后受體鼠分成2組。1、空白對(duì)照組經(jīng)尾靜脈注射生理鹽水；2、試驗(yàn)組經(jīng)尾靜脈注射第三方大鼠臍帶間充質(zhì)干細(xì)胞；結(jié)果顯示,移植心臟的存活時(shí)間為組1：7.5±0.5天；組2：14.8±2.1天。統(tǒng)計(jì)學(xué)分析顯示,移植心臟存活時(shí)間組2比組1明顯延長(zhǎng)(p0.05)。 本研究結(jié)果表明,采用消化法和組織塊培養(yǎng)法均能夠從大鼠的臍帶中獲得間充質(zhì)干細(xì)胞,消化法的效果更佳,臍帶源間充質(zhì)干細(xì)胞作為第三方細(xì)胞,能夠在體外抑制淋巴細(xì)胞的增殖,并能夠在體內(nèi)延長(zhǎng)同種異體心臟移植物的存活時(shí)間和減輕排斥反應(yīng)。
[Abstract]:Mesenchymal stem cell (MScs) is a kind of pluripotent stem cells derived from mesoderm in early stage of development. At present, MSCs mainly comes from adult bone marrow, but many studies show that human umbilical cord is an ideal source of MSCs. There are many studies on rat bone marrow-derived MSCs for transplantation of immune tolerance and tissue repair. However, there is no related research on MSCs derived from rat umbilical cord, so the MSCs derived from umbilical cord can not be tested in vivo. In this study, mesenchymal stem cells (MSCs) were isolated from rat umbilical cord and their biological characteristics were studied. The suitable isolation and culture methods were found, and in vitro and in vivo experiments were carried out to explore the reduction of immune rejection by umbilical cord derived mesenchymal stem cells. The effect of inducing immune tolerance. In the first part of this study, collagenase digestion and tissue mass culture were used to obtain mesenchymal stem cells derived from rat umbilical cord. The results showed that the adherent cells obtained by both methods were mesenchymal stem cells, and their biological characteristics were close to those of bone marrow mesenchymal stem cells reported in the literature. Collagenase digestion was more efficient than tissue culture. In the second part, the spleen cells of Lewis rats were used as reaction cells, and the spleen cells of DA rats were used as stimulating cells. The mesenchymal stem cells derived from the umbilical cord of Wistar rats were added to the mesenchymal stem cells. The mixed lymphocytes were cultured by 3H-thymidine incorporation. The effects of mesenchymal stem cells derived from rat bone umbilical cord on the culture of mixed lymphocytes in vitro were detected. The results showed that mesenchymal stem cells derived from umbilical cord did not induce lymphocyte proliferation in rats, but had significant inhibitory effects on mixed lymphocyte culture in rats. The third part consisted of DA rats as donor and Lewis rats as receptors. A rat model of heterotopic heart transplantation was established and mesenchymal stem cells (MSCs) derived from umbilical cord of Wistar rats were injected into caudal vein immediately after operation to investigate the effect of mesenchymal stem cells (MSCs) on allogeneic heart transplantation in rats. The recipient mice were divided into two groups: 1, the blank control group was injected with normal saline via tail vein; 2, the third party umbilical cord mesenchymal stem cells were injected into the experimental group through tail vein; the results showed that the survival time of the transplanted heart was 1: 7.5 鹵0.5 days. Group 2: 14.8 鹵2.1 days. Statistical analysis showed that the survival time of transplanted heart was significantly longer in group 2 than in group 1 (p0.05). The results showed that mesenchymal stem cells could be obtained from the umbilical cord of rats by digestion and tissue mass culture, and the effect of digestion was better. Mesenchymal stem cells derived from umbilical cord were used as third party cells. It can inhibit the proliferation of lymphocytes in vitro, prolong the survival time of cardiac allografts and alleviate the rejection in vivo.
【學(xué)位授予單位】：中國(guó)人民解放軍軍醫(yī)進(jìn)修學(xué)院
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2009
【分類號(hào)】：R392

【參考文獻(xiàn)】

相關(guān)期刊論文 前10條

1 莊聰文,翁向群,黃盛東,李莉,于偉勇,徐志云;大鼠頸部異位心臟移植急性排斥反應(yīng)的早期病理變化[J];東南國(guó)防醫(yī)藥;2005年03期

2 呂璐璐;劉擁軍;許貞書(shū);王彤;張浪輝;朱雄鵬;梁琳慧;王晗;陳志哲;韓忠朝;;臍帶源間充質(zhì)干細(xì)胞的分離和生物學(xué)性狀[J];福建醫(yī)科大學(xué)學(xué)報(bào);2006年02期

3 張慶殷,郝潔,郭宏偉,劉家望,謝蜀生;門(mén)靜脈輸注供體脾細(xì)胞誘導(dǎo)異基因大鼠心臟移植耐受[J];免疫學(xué)雜志;2001年04期

4 王躍春;張洹;;人胚胎骨髓間充質(zhì)干細(xì)胞的增殖能力及其向三個(gè)胚層起源細(xì)胞分化的特性[J];生理學(xué)報(bào);2008年03期

5 陳健琳,郭子寬,徐晨,李欲航,侯春梅,毛寧,陳虎;間充質(zhì)干細(xì)胞分泌TGF-β1抑制異體T細(xì)胞反應(yīng)性[J];中國(guó)實(shí)驗(yàn)血液學(xué)雜志;2002年04期

6 何津,張毅,江小霞,劉剛,劉元林,李荷蓮,毛寧;人骨髓間充質(zhì)干細(xì)胞對(duì)臍血T淋巴細(xì)胞轉(zhuǎn)化的影響[J];中國(guó)實(shí)驗(yàn)血液學(xué)雜志;2003年01期

7 邱志勇;楊少光;陳振平;趙欽軍;陳曉梨;周澤平;楊仁池;韓忠朝;;臍帶組織源間充質(zhì)干細(xì)胞調(diào)控ITP患者脾細(xì)胞釋放抗血小板抗體的體外研究[J];中國(guó)實(shí)驗(yàn)血液學(xué)雜志;2008年06期

8 同志超;楊鎮(zhèn);同志勤;王坤正;楊團(tuán)民;石宗利;李毅;郭雄;;骨髓間質(zhì)干細(xì)胞體外定向誘導(dǎo)分化為軟骨細(xì)胞的實(shí)驗(yàn)研究[J];中國(guó)骨傷;2008年05期

9 郭子寬,楊靖清,劉曉丹,李秀森,侯春梅,唐佩弦,毛寧;骨髓間葉干細(xì)胞的生物學(xué)特性(英文)[J];Chinese Medical Journal;2001年09期

10 唐偉,馬建波,張?zhí)杖?張慶殷,劉家望,謝蜀生;供體骨髓細(xì)胞輸注促進(jìn)小鼠皮膚移植耐受及其機(jī)理的研究[J];中華微生物學(xué)和免疫學(xué)雜志;2003年02期

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