發(fā)布時間：2018-03-14 13:09

  本文選題：汗腺樣細胞　切入點：骨髓間充質(zhì)干細胞　出處：《內(nèi)蒙古大學(xué)》2017年碩士論文　論文類型：學(xué)位論文

【摘要】：皮膚是機體最大的器官,作為機體最表層,皮膚像一道屏障覆于機體的表面,起到保持皮膚水分,維持正常生理功能的作用。當(dāng)皮膚受到外界損傷或者疾病等因素的影響而造成缺損時,會對生命體造成嚴(yán)重的危害,甚至危及生命。研究發(fā)現(xiàn),細胞治療有助于皮膚損傷的修復(fù)。本研究主要探討由大鼠骨髓間充質(zhì)干細胞(Rat Bone marrow mesenchymal stem cells,rBM-MSCs)誘導(dǎo)的汗腺樣細胞(sweat gland-like cells)與羊脫細胞真皮基質(zhì)(acellular dermal matrix,ADM)復(fù)合后對創(chuàng)面愈合的影響。首先,本研究以大鼠骨髓為材料來源,采用全骨髓貼壁法培養(yǎng)獲得目的細胞。通過形態(tài)、增殖能力和表面標(biāo)志的鑒定顯示出所得細胞為rBM-MSCs。其次,通過將rBM-MSCs與大鼠熱休克汗腺細胞(sweat glands cells,SGCs)共培養(yǎng)的方法進行細胞誘導(dǎo)。RT-PCR和細胞免疫熒光檢測顯示:誘導(dǎo)所得細胞為汗腺樣細胞。隨后,采用酶消化法制備出羊ADM,通過MTS法檢測細胞毒性和HE染色檢測其脫細胞程度顯示:ADM脫細胞程度較為徹底,且沒有細胞毒性。最后,分別將復(fù)合有汗腺樣細胞的ADM和未復(fù)合汗腺樣細胞的ADM覆蓋于裸鼠損傷創(chuàng)面上。通過對創(chuàng)面愈合率、參與創(chuàng)面愈合的蛋白VEGF和Collagen-Ⅰ的表達情況、愈合創(chuàng)面細胞凋亡相關(guān)基因Bax/Bcl-2的相對表達量的變化的測定顯示:汗腺樣細胞復(fù)合羊ADM抑制了愈合創(chuàng)面組織的細胞凋亡并且加快了創(chuàng)面的愈合速率。綜上所述,由BM-MSCs誘導(dǎo)分化的汗腺樣細胞與羊ADM復(fù)合后抑制了愈合創(chuàng)面組織的細胞凋亡并且加快了創(chuàng)面的愈合速率。本研究初步探討了大鼠汗腺樣細胞復(fù)合羊ADM對于創(chuàng)面愈合的影響,為后續(xù)深入研究其作用機制提供了一定的依據(jù)。
[Abstract]:The skin is the largest organ of the body. As the most superficial part of the body, the skin is like a barrier covering the surface of the body, which helps to maintain the moisture of the skin. The function of maintaining normal physiological function. When the skin is damaged by external injury or disease and other factors, it will cause serious harm to the life body, and even endanger the life. The purpose of this study was to investigate the effects of Bone marrow mesenchymal stem cells (rBM-MSCs) induced by rat bone marrow mesenchymal stem cells (Bone marrow mesenchymal stem cells rBM-MSCs) on wound healing after the combination of sweat gland-like cells and sheep acellular dermal matrix adm. In this study, the target cells were obtained from rat bone marrow by whole bone marrow adherent method. The morphology, proliferative ability and surface markers showed that the cells were rBM-MSCs. By co-culture of rBM-MSCs and sweat glands cells SGCsof rat sweat gland cells under heat shock, the results of cell induction. RT-PCR and immunofluorescence analysis showed that the cells induced were sweat gland like cells. Sheep ADMs were prepared by enzyme digestion. The cytotoxicity was detected by MTS method and the degree of acellular by HE staining showed that the acellular degree of ADM was relatively thorough and had no cytotoxicity. The ADM and ADM of the combined sweat gland like cells and the uncomplex sweat gland like cells were respectively covered on the injured wound of nude mice. The expression of protein VEGF and Collagen- 鈪，

本文編號：1611345