本文選題：人臍帶間充質(zhì)干細(xì)胞　切入點(diǎn)：細(xì)胞分化　出處：《內(nèi)蒙古農(nóng)業(yè)大學(xué)》2017年碩士論文　論文類型：學(xué)位論文

【摘要】：角質(zhì)形成細(xì)胞是皮膚表皮組織中最主要的細(xì)胞類型,也是對(duì)皮膚損傷進(jìn)行細(xì)胞治療時(shí)最重要的細(xì)胞類型。間充質(zhì)干細(xì)胞可經(jīng)誘導(dǎo)分化為多種細(xì)胞,具有極低的免疫原性,在細(xì)胞治療領(lǐng)域是一種理想的種子細(xì)胞。本研究通過(guò)改良的"組織塊二次貼壁法",從臍帶華通氏膠中高效、大量地獲得了人臍帶間充質(zhì)干細(xì)胞(human umbilical cord mesenchymal stem cells,hUCMSCs),在特殊的誘導(dǎo)培養(yǎng)基中培養(yǎng)時(shí)可分化為多種細(xì)胞類型。主要研究結(jié)果如下:1.建立了 hUCMSCs細(xì)胞系,細(xì)胞形態(tài)為長(zhǎng)梭形、呈漩渦狀生長(zhǎng)。第三代(P3)細(xì)胞表達(dá)干細(xì)胞轉(zhuǎn)錄因子Sox2、間充質(zhì)干細(xì)胞表面標(biāo)志物基因CD29、CD90和CD105,不表達(dá)造血系細(xì)胞表面標(biāo)志物基因CD34、CD45及人白細(xì)胞抗原基因HLA-DR。經(jīng)測(cè)定,其生長(zhǎng)曲線呈"S"型,細(xì)胞倍增時(shí)間為24.7 h,生長(zhǎng)良好、活性很高,染色體為46,XY正常男性核型,遺傳穩(wěn)定。2.建立了 hUCMSCs多向誘導(dǎo)分化培養(yǎng)體系,誘導(dǎo)其向外胚層(神經(jīng)細(xì)胞、角質(zhì)形成細(xì)胞)和中胚層(成骨細(xì)胞)來(lái)源的細(xì)胞分化,染色鑒定結(jié)果均為陽(yáng)性。3.建立了角質(zhì)形成細(xì)胞誘導(dǎo)分化培養(yǎng)體系,hUCMSCs經(jīng)誘導(dǎo)培養(yǎng)后分化為角質(zhì)形成細(xì)胞,蛋白質(zhì)水平染色結(jié)果顯示其表達(dá)細(xì)胞角蛋白,基因水平檢測(cè)到其特異基因CK19的表達(dá)量逐日增高。綜上所述,本實(shí)驗(yàn)創(chuàng)建了 hUCMSCs原代培養(yǎng)的新方法,并成功建立了 hUCMSCs細(xì)胞系,獲得的hUCMSCs具有多向分化的能力,同時(shí)建立了向角質(zhì)形成細(xì)胞誘導(dǎo)分化的培養(yǎng)體系,為間充質(zhì)干細(xì)胞的臨床應(yīng)用尤其是大面積皮膚損傷的細(xì)胞治療提供了實(shí)驗(yàn)依據(jù)。
[Abstract]:Keratinocytes are the most important cell types in skin epidermis and the most important cell types in the treatment of skin injury. Mesenchymal stem cells can be induced to differentiate into a variety of cells with very low immunogenicity. It is an ideal seed cell in the field of cell therapy. A large number of human umbilical cord mesenchymal stem cells were obtained from human umbilical cord mesenchymal stem cells, which could differentiate into many cell types in a special induction medium. The main results are as follows: 1. The cell line of hUCMSCs was established, and the cell morphology was fusiform. The third generation of P3 cells expressed stem cell transcription factor Sox2, mesenchymal stem cell surface marker gene CD29, CD90 and CD105, but not hematopoietic cell surface marker gene CD34, CD45 and human leukocyte antigen gene HLA-DR. Its growth curve is "S" type, the cell doubling time is 24.7 h, the cell growth is good, the activity is very high, the chromosome is 46 XY normal male karyotype, the heredity is stable .2.It has established the culture system of hUCMSCs multidirectional induction differentiation, induced it to the ectoderm (nerve cell), The differentiation of keratinocytes from keratinocytes) and mesoderm (osteoblasts) were positive. 3. The culture system of keratinocyte induced differentiation was established and then differentiated into keratinocytes. The results of protein level staining showed that it expressed cytokeratin, and the expression of its specific gene CK19 was increased day by day. In conclusion, a new method of primary culture of hUCMSCs was established, and the hUCMSCs cell line was successfully established. The obtained hUCMSCs has the ability of multi-differentiation, and the culture system of inducing differentiation into keratinocytes has been established, which provides the experimental basis for the clinical application of mesenchymal stem cells, especially for the treatment of extensive skin injury.
【學(xué)位授予單位】：內(nèi)蒙古農(nóng)業(yè)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R329.2

【參考文獻(xiàn)】

相關(guān)期刊論文 前10條

1 張惟;陳松;王繼明;段紅濤;孔佳慧;王月欣;董蒙;畢雪;宋建;;人臍帶間充質(zhì)干細(xì)胞向神經(jīng)細(xì)胞分化的實(shí)驗(yàn)研究[J];臨床眼科雜志;2015年04期

2 何潔;趙晶;王金祥;蔡學(xué)敏;龐榮清;潘興華;;多次貼壁法高效制備人臍帶間充質(zhì)干細(xì)胞[J];西南國(guó)防醫(yī)藥;2015年08期

3 高彥琳;張寧坤;陳厚良;高連如;朱智明;;改良原代培養(yǎng)體系提高人臍帶間充質(zhì)干細(xì)胞的產(chǎn)量[J];中國(guó)組織工程研究;2015年10期

4 李艷琪;王洪一;姚堯;劉晶晶;徐瀟;張宇;劉洋;吳祖澤;靳繼德;;人臍帶源間充質(zhì)干細(xì)胞分離培養(yǎng)方法的改進(jìn)[J];中國(guó)組織工程研究;2014年10期

5 胡培;王小莉;李東升;嚴(yán)世榮;陳玲;楊曉文;;人臍帶間充質(zhì)干細(xì)胞的分離培養(yǎng)與鑒定[J];生物技術(shù)通訊;2014年01期

6 崔冬冰;何志旭;李永念;王鳳昌;;臍帶間充質(zhì)干細(xì)胞體外傳代培養(yǎng)后細(xì)胞核型穩(wěn)定性[J];貴陽(yáng)醫(yī)學(xué)院學(xué)報(bào);2013年05期

7 徐小紅;王力;張寧坤;鄭楠;高連如;朱智明;;Apelin-13對(duì)5-Aza誘導(dǎo)臍帶間充質(zhì)干細(xì)胞向心肌細(xì)胞分化的影響[J];解放軍醫(yī)學(xué)雜志;2013年10期

8 龐榮清;何潔;李福兵;趙晶;阮光萍;王桂華;瞿良;李黎;蔡學(xué)敏;潘興華;;一種簡(jiǎn)單的人臍帶間充質(zhì)干細(xì)胞分離培養(yǎng)方法[J];中華細(xì)胞與干細(xì)胞雜志(電子版);2011年02期

9 馬錫慧;馮凱;石炳毅;;人臍帶間充質(zhì)干細(xì)胞生物學(xué)特性及其研究進(jìn)展[J];中國(guó)組織工程研究與臨床康復(fù);2011年32期

10 齊凱;董麗媛;陳顯久;趙婕;薛國(guó)芳;陳彥;牛勃;;人臍帶來(lái)源間充質(zhì)干細(xì)胞分離培養(yǎng)方法的優(yōu)化[J];中國(guó)組織工程研究與臨床康復(fù);2011年23期

，

本文編號(hào)：1600391