本文選題：干細(xì)胞 + 脂肪干細(xì)胞��； 參考：《中國(guó)組織工程研究》2014年10期

【摘要】：背景:如何運(yùn)用無血清培養(yǎng)體系大規(guī)模擴(kuò)增處于未分化狀態(tài)的脂肪間充質(zhì)干細(xì)胞,并使其保持"干性"是脂肪間充質(zhì)干細(xì)胞臨床應(yīng)用轉(zhuǎn)化中急待解決的難題。目的:建立含細(xì)胞外基質(zhì)的脂肪間充質(zhì)干細(xì)胞體外培養(yǎng)系統(tǒng),驗(yàn)證其擴(kuò)增細(xì)胞的高效性、有效性及安全性。方法:在無血清培養(yǎng)條件下,將體外分離得到的脂肪間充質(zhì)干細(xì)胞分別接種在包被細(xì)胞外基質(zhì)的培養(yǎng)板和傳統(tǒng)二維塑料培養(yǎng)板上。經(jīng)過體外擴(kuò)增后,比較2種條件下細(xì)胞數(shù)量、細(xì)胞表面標(biāo)記物表達(dá)、細(xì)胞衰老狀況以及體外多向分化能力(誘導(dǎo)成脂、成骨和成軟骨)的差異,考察脂肪間充質(zhì)干細(xì)胞在含細(xì)胞外基質(zhì)的培養(yǎng)條件下擴(kuò)增后的臨床安全性。結(jié)果與結(jié)論:脂肪間充質(zhì)干細(xì)胞擴(kuò)增到第5代時(shí),包被細(xì)胞外基質(zhì)培養(yǎng)板的細(xì)胞產(chǎn)量已經(jīng)是傳統(tǒng)二維塑料培養(yǎng)板的10倍以上,流式細(xì)胞檢測(cè)表明,在含細(xì)胞外基質(zhì)條件下擴(kuò)增的脂肪間充質(zhì)干細(xì)胞仍保持了干細(xì)胞表面特定標(biāo)記物的表達(dá)。細(xì)胞衰老檢測(cè)結(jié)果顯示,使用包被細(xì)胞外基質(zhì)的培養(yǎng)板擴(kuò)增脂肪間充質(zhì)干細(xì)胞至第15代時(shí),細(xì)胞仍幾乎無老化現(xiàn)象,而使用二維塑料培養(yǎng)板擴(kuò)增的細(xì)胞在第5代時(shí)已出現(xiàn)明顯老化,傳代后細(xì)胞增殖能力明顯下降。體外多向誘導(dǎo)分化實(shí)驗(yàn)顯示,脂肪間充質(zhì)干細(xì)胞在含細(xì)胞外基質(zhì)條件下擴(kuò)增至第15代時(shí)仍具有成脂、成骨和成軟骨的分化功能,并且與第5代時(shí)沒有明顯差異,同時(shí)顯著優(yōu)于傳統(tǒng)培養(yǎng)條件下的第5代脂肪間充質(zhì)干細(xì)胞。染色體核型分析及小鼠成瘤性試驗(yàn)結(jié)果表明,脂肪間充質(zhì)干細(xì)胞在含細(xì)胞外基質(zhì)條件下擴(kuò)增后仍具備臨床應(yīng)用的安全性。以上結(jié)果證實(shí),含細(xì)胞外基質(zhì)的無血清培養(yǎng)系統(tǒng)可以更高效且安全地?cái)U(kuò)增出具有臨床應(yīng)用潛能的脂肪間充質(zhì)干細(xì)胞。
[Abstract]:Background: it is an urgent problem for the clinical application and transformation of adipose mesenchymal stem cells to expand the undifferentiated adipose mesenchymal stem cells in a large scale and keep them dry by using serum-free culture system. Aim: to establish an in vitro culture system of adipose mesenchymal stem cells containing extracellular matrix to verify the efficiency, efficacy and safety of expanded cells. Methods: the adipose mesenchymal stem cells isolated in vitro were inoculated on the culture plate coated with extracellular matrix and on the traditional two-dimensional plastic culture plate under the condition of serum-free culture. After in vitro amplification, the difference of cell number, expression of cell surface markers, cell senescence and the ability of differentiation in vitro (adipogenesis, osteogenesis and cartilage formation) were compared. To investigate the clinical safety of adipose mesenchymal stem cells expanded in extracellular matrix culture. Results and conclusion: when adipose mesenchymal stem cells were expanded to the fifth passage, the cell production of the coated extracellular matrix culture plate was 10 times higher than that of the traditional two-dimensional plastic culture plate. Adipose mesenchymal stem cells containing extracellular matrix maintained the expression of specific markers on the surface of stem cells. The results of cell senescence test showed that the adipose mesenchymal stem cells were expanded with the culture plate coated with extracellular matrix until the 15th generation, the cells still had almost no aging phenomenon. However, the cells amplified by two-dimensional plastic culture plate had obviously aged at the fifth passage, and the proliferation ability of the cells decreased obviously after passage. The multidirectional differentiation experiment in vitro showed that adipose mesenchymal stem cells still had the functions of adipogenesis, osteogenesis and cartilage differentiation at the 15th generation after expansion under the condition of extracellular matrix, and there was no significant difference between adipose mesenchymal stem cells and the fifth generation of adipose mesenchymal stem cells. At the same time, it is superior to the fifth generation adipose mesenchymal stem cells in traditional culture. The results of chromosome karyotype analysis and mouse tumorigenicity test showed that adipose mesenchymal stem cells were still safe for clinical application after expansion under extracellular matrix. These results suggest that the serum-free culture system containing extracellular matrix can effectively and safely amplify adipose mesenchymal stem cells with clinical application potential.
【作者單位】： 解放軍第455醫(yī)院 解放軍南京軍區(qū)生物細(xì)胞工程技術(shù)醫(yī)學(xué)轉(zhuǎn)化基地干細(xì)胞實(shí)驗(yàn)室;上海交通大學(xué)醫(yī)學(xué)院附屬仁濟(jì)醫(yī)院消化科 上海市消化疾病研究所干細(xì)胞實(shí)驗(yàn)室;
【基金】：上海市科委浦江人才計(jì)劃項(xiàng)目(09PJ1407300) 國(guó)家自然科學(xué)基金項(xiàng)目(30971468)~~
【分類號(hào)】：R329

【共引文獻(xiàn)】

相關(guān)期刊論文 前10條

1 Brian E. Grottkau;Xingmei Yang;Liang Zhang;Ling Ye;Yunfeng Lin;;Comparison of Effects of Mechanical Stretching on Osteogenic Potential of ASCs and BMSCs[J];Bone Research;2013年03期

2 王波;唐曉鵬;;臍血干細(xì)胞與骨髓干細(xì)胞聯(lián)合移植治療大鼠急性肝衰竭實(shí)驗(yàn)研究[J];實(shí)用肝臟病雜志;2014年01期

3 劉真真;李海洋;鄧彥飛;喬樹葉;朱鵬;楊素芳;石德順;;家畜脂肪干細(xì)胞研究進(jìn)展[J];基因組學(xué)與應(yīng)用生物學(xué);2014年01期

4 趙佳佳;胡麗;劉加榮;宮妮雅;陳莉莉;;脂肪干細(xì)胞來源的生長(zhǎng)因子與口腔黏膜成纖維細(xì)胞增殖[J];中國(guó)組織工程研究;2013年32期

5 郭\厴，

本文編號(hào)：1945243