骨髓间充质干细胞与内皮细胞平面共培养促进微血管网形成的初步研究

发布时间：2018-03-08 16:25

  本文选题：骨髓间充质干细胞　切入点：内皮细胞　出处：《组织工程与重建外科杂志》2016年05期 　论文类型：期刊论文

【摘要】：目的研究大鼠骨髓间充质干细胞(BMSCs)与内皮细胞(ECs)在不使用Matrigel等基质,以及在含或不含血清的条件下,于普通平面培养皿上共培养的成网能力,为日后两类细胞相互作用的研究提供一种外界干扰最少的共培养条件。方法以全骨髓贴壁培养法分离培养大鼠BMSCs,并从培养形态、细胞表面标记物和三系分化能力等方面予以鉴定。以添加或不添加10%FBS的低糖DMEM为培养液,将大鼠BMSCs与ECs共培养于普通平面培养皿上,连续观察微血管网的形成过程,并通过CM-Dil标记特定细胞的方法了解血管网中细胞构成。平滑肌细胞标记物SM22α,Calponin及内皮细胞标记物CD31被用于检测上述条件下微血管网中细胞标记物变化。结果通过全骨髓贴壁培养可分离并获得大鼠BMSCs。在含10%FBS条件下,细胞于普通平面培养皿上共培养8 d可形成明显的微血管网,微血管网由大鼠BMSCs和ECs共同构成;在无血清共培养条件下,大鼠BMSCs和ECs可于24 h内共同参与形成微血管网。免疫荧光染色显示CD31+细胞和SM22α+/Calponin+细胞共同构成网状结构。结论在普通平面培养皿上,以含血清或不含血清的培养液共培养大鼠BMSCs和ECs均可形成微血管网。
[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 伪 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 伪 -Calponin cells formed a network structure. Both BMSCs and ECs co-cultured in serum-containing or serum-free medium could form microvascular networks.
【作者单位】： 上海交通大学医学院附属上海儿童医学中心心胸外科;上海交通大学医学院附属上海儿童医学中心儿科转化医学研究所;
【基金】：国家自然科学基金(31200735) 上海市科委基金(134119a0400,15411966800) 上海卫生和计划生育委员会项目(20144Y0166) 东南大学生物电子学国家重点实验室开放研究基金
【分类号】：R329.2
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