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热休克内皮细胞诱导骨髓间充质干细胞向血管内皮细胞分化
引用本文:曹百川,曾高峰,高云兵,邓贵营,岑忠喜,张传阳,郭彦德,宗少晖. 热休克内皮细胞诱导骨髓间充质干细胞向血管内皮细胞分化[J]. 中国组织工程研究, 2020, 24(19): 3023-3028. DOI: 10.3969/j.issn.2095-4344.2081
作者姓名:曹百川  曾高峰  高云兵  邓贵营  岑忠喜  张传阳  郭彦德  宗少晖
作者单位:广西医科大学第一附属医院脊柱骨病外科,广西壮族自治区南宁市 530021;广西医科大学公共卫生学院,广西壮族自治区南宁市 530021
基金项目:广西重点研发计划(桂科AB17195001),项目负责人:曾高峰;广西高等学校高水平创新团队及卓越学者计划,项目负责人:宗少晖;广西医学高层次骨干人才“139”计划,项目负责人:宗少晖~~
摘    要:文题释义:血管内皮细胞:研究中一般称内皮细胞,通常指衬于心、血管和淋巴管内表面的单层扁平上皮,它形成血管的内壁。它们具有吞噬异物、细菌、坏死和衰老组织的功能,还参与机体免疫活动功能。热休克:组织工程中一种细胞处理方法,将细胞置于42 ℃(也有文献报道为47 ℃)中1 h,造成热休克状态。背景:目前关于间充质干细胞向内皮细胞分化的研究,多采用细胞因子或2种细胞共培养的方法进行诱导,热休克处理的内皮细胞诱导间充质干细胞向内皮细胞分化尚未见报道。目的:观察热休克处理的人脐静脉内皮细胞诱导骨髓间充质干细胞向血管内皮细胞分化的能力,并探究诱导骨髓间充质干细胞形成血管的能力。方法:将热休克处理后的人脐静脉内皮细胞与人骨髓间充质干细胞体外非接触共培养,诱导14 d后采用流式细胞仪和免疫荧光检测骨髓间充质干细胞中CD144、CD31、VEGFR2、vWF表型的表达;将诱导14 d后的骨髓间充质干细胞、未诱导的骨髓间充质干细胞移植到裸鼠皮下,14 d后取移植物做苏木精-伊红染色,观察体内血管形成能力;Matrigel成血管实验观察诱导14 d后的骨髓间充质干细胞和未诱导的骨髓间充质干细胞的体外血管生成能力。结果与结论:①共培养后骨髓间充质干细胞形态改变,呈类似铺路石状排列,流式细胞仪及细胞免疫荧光结果显示共培养后骨髓间充质干细胞VEGFR2、CD31、CD144、vWF表达增加;②体内移植物苏木精-伊红染色显示诱导后的骨髓间充质干细胞排列较对照组规律,有成血管倾向;③体外Matrigel成血管实验显示诱导后骨髓间充质干细胞成血管能力较对照组有所增加;④结果表明,与热休克处理的人脐静脉内皮细胞共培养能促进人骨髓间充质干细胞向内皮细胞分化,内皮细胞特异性表型转化较明显,具有一定血管形成倾向。ORCID: 0000-0003-4089-8882(曹百川)中国组织工程研究杂志出版内容重点:干细胞;骨髓干细胞;造血干细胞;脂肪干细胞;肿瘤干细胞;胚胎干细胞;脐带脐血干细胞;干细胞诱导;干细胞分化;组织工程

关 键 词:骨髓间充质干细胞  内皮细胞  热休克  共培养  分化  血管生成  
收稿时间:2019-11-30

Heat-shock endothelial cells induce differentiation of bone marrow mesenchymal stem cells into vascular endothelial cells
Cao Baichuan,Zeng Gaofeng,Gao Yunbing,Deng Guiying,Cen Zhongxi,Zhang Chuanyang,Guo Yande,Zong Shaohui. Heat-shock endothelial cells induce differentiation of bone marrow mesenchymal stem cells into vascular endothelial cells[J]. Chinese Journal of Tissue Engineering Research, 2020, 24(19): 3023-3028. DOI: 10.3969/j.issn.2095-4344.2081
Authors:Cao Baichuan  Zeng Gaofeng  Gao Yunbing  Deng Guiying  Cen Zhongxi  Zhang Chuanyang  Guo Yande  Zong Shaohui
Affiliation:Departmentof Spine Osteopathia, the First Affiliated Hospital of Guangxi MedicalUniversity, Nanning 530021, Guangxi Zhuang Autonomous Region, China; Schoolof Public Health, Guangxi Medical University, Nanning 530021, Guangxi ZhuangAutonomous Region, China
Abstract:BACKGROUND:Current research on the differentiation of mesenchymal stem cells into endothelial cells mainly focuses on the induction of cytokines or co-culture of two types of cells.Research has been not yet reported on heat shock-treated endothelial cells inducing the differentiation of mesenchymal stem cells into endothelial cells.OBJECTIVE:To investigate the ability of heat shock-treated endothelial cells to induce bone marrow mesenchymal stem cells to differentiate into vascular endothelial cells,and to explore the angiogenic capacity of induced bone marrow mesenchymal stem cells.METHODS:Human bone marrow mesenchymal stem cells were co-cultured with heat shock-treated human umbilical vein endothelial cells in a non-contact manner.The expression of CD31,CD144,VEGFR2 and vWF in the bone marrow mesenchymal stem cells was detected by flow cytometry at 14 days after co-culture.The immunofluorescence of the cells was further verified.In vivo angiogenesis test was detected using hematoxylin-eosin staining in nude mice undergoing subcutaneous transplantation of induced and non-induced bone marrow mesenchymal stem cells.Matrigel angiogenesis test was applied to observe the angiogenic ability of induced and non-induced bone marrow mesenchymal stem cells.RESULTS AND CONCLUSION:After coculture,bone marrow mesenchymal stem cells had paving-stone like structure.Flow cytometry and cellular immunofluorescence results showed an increase in the expression of VEGFR2,vWF,CD31 and CD144 after co-culture.The hematoxylin-eosin staining of the graft showed that the induced bone marrow mesenchymal stem cells arranged in a regular manner and had an angiogenic tendency.The Matrigel angiogenesis test showed that the angiogenic ability of bone marrow mesenchymal stem cells was increased after induction compared with the control group.These findings indicate that co-culture with heat shock-treated human umbilical vein endothelial cells can promote the differentiation of mesenchymal stem cells into endothelial cells,and the specific phenotype of endothelial cells is obviously transformed,which has a certain tendency of angiogenesis.
Keywords:bone marrow mesenchymal stem cells  endothelial cells  heat shock  co-culture  differentiation  angiogenesis
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