胶原海绵促进人绒毛膜来源间充质干细胞向血管内皮细胞分化的研究
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摘要
探索人绒毛膜来源间充质干细胞与胶原海绵的生物相容性及复合培养向血管内皮细胞分化。应用IV型胶原酶消化的方法,从胎盘组织中分离人绒毛膜间充质细胞,培养第2代细胞,进行内皮诱导后,种植于Matrigel基质胶并观察成管情况;细胞与胶原海绵复合培养,通过HE染色以及免疫荧光染色,鉴定CD31和vWF的表达。人绒毛膜间充质干细胞向血管内皮定向诱导2周后,形态由长梭形、旋涡状生长逐步变为圆形、铺路石状,免疫荧光染色结果显示CD31和vWF阳性表达;种植于Matrigel胶12 h后表现出成管功能,与胶原海绵复合培养,经HE染色,可见细胞分布于材料内部,免疫荧光染色可见CD31和vWF阳性表达细胞。人绒毛膜间充质干细胞具有定向诱导分化为血管内皮细胞的潜能,胶原海绵具有良好的生物相容性,可以作为组织工程研究的支架材料。
To explore the feasibility of isolation and culture mesenchymal stem cells from human chorionic membrane and the biocompatibility with collagen sponge, also the ability to differentiation into endothelial cells in vitro or co-culture with collagen sponge. Using type IV collagenase digestion method,the mesenchymal stem cells of chorionic membrane from human placenta was isolated and cultured. The second-generation culture cells were used to induction the endothelial cells and planted on Matrigel gel to observe the vascular function. To analyze the biocompatibility of cells and biomaterials, the expression of CD31 and vWF were identified by immunofluorescence staining after the induced cells planted on collagen sponge. The morphology by long fusiform, spiral growth of human chorionic mesenchymal stem cells gradually became round, paving stone like two weeks after directional induced into vascular endothelial cells. Immunofluorescence staining showed the induced cells positive expression of CD31 and vWF. HE staining showed that the cells were distributed in the collagen sponge and CD31 and vWF positive cells were found by immunofluorescence staining. The human chorionic membrane mesenchymal stem cells have the potential of differentiation into vascular endothelial cells and collagen sponge has good biocompatibility, which can be used for tissue engineering research.
To explore the feasibility of isolation and culture mesenchymal stem cells from human chorionic membrane and the biocompatibility with collagen sponge, also the ability to differentiation into endothelial cells in vitro or co-culture with collagen sponge. Using type IV collagenase digestion method,the mesenchymal stem cells of chorionic membrane from human placenta was isolated and cultured. The second-generation culture cells were used to induction the endothelial cells and planted on Matrigel gel to observe the vascular function. To analyze the biocompatibility of cells and biomaterials, the expression of CD31 and vWF were identified by immunofluorescence staining after the induced cells planted on collagen sponge. The morphology by long fusiform, spiral growth of human chorionic mesenchymal stem cells gradually became round, paving stone like two weeks after directional induced into vascular endothelial cells. Immunofluorescence staining showed the induced cells positive expression of CD31 and vWF. HE staining showed that the cells were distributed in the collagen sponge and CD31 and vWF positive cells were found by immunofluorescence staining. The human chorionic membrane mesenchymal stem cells have the potential of differentiation into vascular endothelial cells and collagen sponge has good biocompatibility, which can be used for tissue engineering research.
引文
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[2]Oswald J,Boxberger S,J?rgensen B,et al.Mesenchymal stem cells can be differentiated into endothelial cells in vitro[J].Stem Cells,2004,22(3):377-384.
[3]Ikhapoh I,Pelham C J,Agrawal D K.Synergistic effect of angiotensin II on vascular endothelial growth factor-A-mediated differentiation of bone marrow-derived mesenchymal stem cells into endothelial cells[J].Stem Cell Res Ther,2015,6(1):4.
[4]Jaramillo-Ferrada P A,Wolvetang E J,Cooper-White J J.Differential mesengenic potential and expression of stem cell-fate modulators in mesenchymal stromal cells from human-term placenta and bone marrow[J].J Cell Physiol,2012,227(9):3234-3342.
[5]Miao Zongning,Sun Hongli,Xue Yifeng.Isolation and characterization of human chorionic membranes mesenchymal stem cells and their neural differentiation[J].Tissue Eng Regen Med,2017,14(2):143-151.
[6]苗宗宁,吴卫江,钱寒光,等.新生大鼠海马神经干细胞的分离培养及分化研究[J].生物医学工程研究,2016,35(4):303-308.
[7]Keating A.Mesenchymal stromal cells:new directions[J].Cell Stem Cell,2012,10(6):709-716.
[8]汪黎明,陈鑫,徐明.组织工程血管支架材料最适孔径的研究[J].中华实验外科杂志,2008,25(2):161-162.
[9]Glowacki J,Mizuno S,Greenberger J S.Perfusion enhances functions of bone marrow stromal cells in three-dimensional culture[J].Cell Transplant,1998,7(3):319-326.
[10]杨逸禧,杨波,娄爱菊,等.关于三维共培养体系中血管化的研究[J].中华实验外科杂志,2017,34(11):1880-1884.