本文選題：人羊水來(lái)源間充質(zhì)干細(xì)胞　切入點(diǎn)：軟骨細(xì)胞　出處：《復(fù)旦大學(xué)》2009年碩士論文

【摘要】： 研究目的 探討分離、提純和擴(kuò)增孕中期羊水來(lái)源間充質(zhì)干細(xì)胞(AF-MSCs)的方法,以及體外誘導(dǎo)AF-MSCs向軟骨細(xì)胞分化的可能性。 研究方法 機(jī)械手段挑取孕中期人羊水細(xì)胞培養(yǎng)體系中的MSCs集落,培養(yǎng)擴(kuò)增后,通過(guò)流式細(xì)胞術(shù)對(duì)表面抗原表達(dá)情況進(jìn)行鑒定,并通過(guò)RT-PCR檢測(cè)多能性基因Oct4及Nanog的表達(dá)情況。取第3代AF-MSCs,經(jīng)TGF-β1、IGF-1、地塞米松等生長(zhǎng)因子向軟骨細(xì)胞方向誘導(dǎo)分化。倒置相差顯微鏡觀察誘導(dǎo)后細(xì)胞形態(tài)的變化。誘導(dǎo)3周后,免疫熒光染色及RT-PCR分別檢測(cè)軟骨特異性基質(zhì)CollengonⅡ、Aggrecan、Chondromudulin-1、S100的蛋白及其基因的表達(dá)情況。 研究結(jié)果 機(jī)械手段分離出的AF-MSCs為CD105、CD166陽(yáng)性,CD29若陽(yáng)性,CD34、CD45陰性,符合MSCs特點(diǎn)。AF-MSCs表達(dá)多能性基因Oct4,但不表達(dá)Nanog。誘導(dǎo)2周后,細(xì)胞開(kāi)始呈多角形樣改變;誘導(dǎo)后3周時(shí),大部分細(xì)胞由長(zhǎng)梭形轉(zhuǎn)變?yōu)槎嘟切�。免疫熒光及RT-PCR檢測(cè)顯示誘導(dǎo)后的AF-MSCs軟骨特異性基質(zhì)CollengonⅡ、Aggrecan、Chondromudulin-1和S100的蛋白和基因呈陽(yáng)性表達(dá),而未經(jīng)誘導(dǎo)培養(yǎng)的AF-MSCs呈陰性表達(dá)。 研究結(jié)論 機(jī)械分離法可有效獲得孕中期羊水來(lái)源間充質(zhì)干細(xì)胞,經(jīng)TGF-β1,IGF-1,地塞米松聯(lián)合誘導(dǎo)3周后,AF-MSCs可以分化為軟骨細(xì)胞樣細(xì)胞。
[Abstract]:Research purpose. To explore the methods of isolation, purification and amplification of AF-MSCs derived from amniotic fluid in the second trimester of pregnancy, and the possibility of inducing AF-MSCs to differentiate into chondrocytes in vitro. Research method. In the second trimester of human amniotic fluid cell culture system, the MSCs colony was selected by mechanical means, and the expression of surface antigen was identified by flow cytometry. The expression of Oct4 and Nanog was detected by RT-PCR. The third generation AF-MSCs were induced to differentiate into chondrocytes by growth factors such as TGF- 尾 1 IGF-1 and dexamethasone. The morphologic changes of the cells were observed by inverted phase contrast microscopy. The expression of Chondromudulin-1 S100 protein and its gene were detected by immunofluorescence staining and RT-PCR, respectively. Research results. The AF-MSCs isolated by mechanical means was CD105, CD166 positive and CD29 positive. If CD34 + CD45 was negative, it was consistent with the characteristics of MSCs. AF-MSCs expressed multipotent gene Oct4, but did not express Nanog.After 2 weeks of induction, the cells began to show polygonal changes, and at 3 weeks after induction, the cells began to show polygonal changes. Immunofluorescence and RT-PCR analysis showed that the AF-MSCs chondromudin-1 and S100 chondromudin-1 and S100 chondromudin-1 were expressed positively, but the uninduced AF-MSCs showed negative expression. Research conclusion. Mesenchymal stem cells derived from amniotic fluid in the second trimester of pregnancy could be obtained by mechanical isolation. AF-MSCs could differentiate into chondrocyte-like cells after induction of TGF- 尾 _ 1 IGF-1 and dexamethasone for 3 weeks.
【學(xué)位授予單位】：復(fù)旦大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2009
【分類號(hào)】：R329

【參考文獻(xiàn)】

相關(guān)期刊論文 前1條

1 Patrizia Bossolasco;Tiziana Montemurro;Lidia Cova;Stefano Zangrossi;Cinzia Calzarossa;Simona Buiatiotis;Davide Soligo;Silvano Bosari;Vincenzo Silani;Giorgio Lambertenghi Deliliers;PaoloRebulla;LorenzaLazzari;;Molecular and phenotypic characterization of human amniotic fluid cells and their differentiation potential[J];Cell Research;2006年04期

，

本文編號(hào)：1659043