本文選題：骨髓間充質(zhì)干細(xì)胞　切入點(diǎn)：內(nèi)皮細(xì)胞　出處：《組織工程與重建外科雜志》2016年05期 　論文類型：期刊論文

【摘要】：目的研究大鼠骨髓間充質(zhì)干細(xì)胞(BMSCs)與內(nèi)皮細(xì)胞(ECs)在不使用Matrigel等基質(zhì),以及在含或不含血清的條件下,于普通平面培養(yǎng)皿上共培養(yǎng)的成網(wǎng)能力,為日后兩類細(xì)胞相互作用的研究提供一種外界干擾最少的共培養(yǎng)條件。方法以全骨髓貼壁培養(yǎng)法分離培養(yǎng)大鼠BMSCs,并從培養(yǎng)形態(tài)、細(xì)胞表面標(biāo)記物和三系分化能力等方面予以鑒定。以添加或不添加10%FBS的低糖DMEM為培養(yǎng)液,將大鼠BMSCs與ECs共培養(yǎng)于普通平面培養(yǎng)皿上,連續(xù)觀察微血管網(wǎng)的形成過程,并通過CM-Dil標(biāo)記特定細(xì)胞的方法了解血管網(wǎng)中細(xì)胞構(gòu)成。平滑肌細(xì)胞標(biāo)記物SM22α,Calponin及內(nèi)皮細(xì)胞標(biāo)記物CD31被用于檢測(cè)上述條件下微血管網(wǎng)中細(xì)胞標(biāo)記物變化。結(jié)果通過全骨髓貼壁培養(yǎng)可分離并獲得大鼠BMSCs。在含10%FBS條件下,細(xì)胞于普通平面培養(yǎng)皿上共培養(yǎng)8 d可形成明顯的微血管網(wǎng),微血管網(wǎng)由大鼠BMSCs和ECs共同構(gòu)成;在無血清共培養(yǎng)條件下,大鼠BMSCs和ECs可于24 h內(nèi)共同參與形成微血管網(wǎng)。免疫熒光染色顯示CD31+細(xì)胞和SM22α+/Calponin+細(xì)胞共同構(gòu)成網(wǎng)狀結(jié)構(gòu)。結(jié)論在普通平面培養(yǎng)皿上,以含血清或不含血清的培養(yǎng)液共培養(yǎng)大鼠BMSCs和ECs均可形成微血管網(wǎng)。
[Abstract]:Objective to study the ability of rat bone marrow mesenchymal stem cells (BMSCs) and endothelial cells (ECs) to co-culture on plain plate without Matrigel and with or without serum. Methods BMSCs of rats were isolated and cultured by whole bone marrow adherent culture method. The cell surface markers and the differentiation ability of three lines were identified. The rat BMSCs and ECs were co-cultured on a flat plate with or without 10 S low-glucose DMEM as the culture medium. The formation of microvascular network was observed continuously. The cell composition in vascular network was studied by CM-Dil labeling method. Smooth muscle cell marker SM22 偽 Calponin and endothelial cell marker CD31 were used to detect the changes of cell markers in microvascular network under the above conditions. Bone marrow adherent culture can be used to isolate and obtain rat BMSCs. The microvascular network was formed by co-culture of cells on common plate culture dish for 8 days. The microvascular network was composed of rat BMSCs and ECs, and was co-cultured in serum-free medium. Rat BMSCs and ECs could participate in the formation of microvascular network within 24 hours. Immunofluorescence staining showed that CD31 cells and SM22 偽 -Calponin cells formed a network structure. Both BMSCs and ECs co-cultured in serum-containing or serum-free medium could form microvascular networks.
【作者單位】： 上海交通大學(xué)醫(yī)學(xué)院附屬上海兒童醫(yī)學(xué)中心心胸外科;上海交通大學(xué)醫(yī)學(xué)院附屬上海兒童醫(yī)學(xué)中心兒科轉(zhuǎn)化醫(yī)學(xué)研究所;
【基金】：國家自然科學(xué)基金(31200735) 上海市科委基金(134119a0400,15411966800) 上海衛(wèi)生和計(jì)劃生育委員會(huì)項(xiàng)目(20144Y0166) 東南大學(xué)生物電子學(xué)國家重點(diǎn)實(shí)驗(yàn)室開放研究基金
【分類號(hào)】：R329.2
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本文編號(hào)：1584671