中国组织工程研究

中国组织工程研究 ›› 2019, Vol. 23 ›› Issue (25): 3951-3955.doi: 10.3969/j.issn.2095-4344.1773

• 骨髓干细胞 bone marrow stem cells • 上一篇    下一篇

骨髓间充质干细胞膜片和脐静脉内皮细胞共培养模型的构建及鉴定

周  震，王亚敏，任  飞，张兆强，曾曙光，杨  熙
  

  1. 南方医科大学口腔医院，广东省广州市  510280
  • 修回日期:2019-04-02 出版日期:2019-09-08 发布日期:2019-09-08
  • 通讯作者: 王亚敏，主治医师，硕士，南方医科大学口腔医院，广东省广州市 510280； 并列通讯作者：任飞，主任医师，南方医科大学口腔医院，广东省广州市 510280
  • 作者简介:周震，男，1983年生，汉族，贵州省人，主治医师，2010年贵州医科大学毕业，硕士。
  • 基金资助:

    广东省自筹经费类科技计划项目(2017ZC0124)，项目负责人：周震；广东省医学科学技术研究基金项目(C2017061)，项目负责人：任飞；广东省自然科学基金项目(2018A030313759)，项目负责人：张兆强；广州市科技计划项目(201707010193)，项目负责人：曾曙光

Co-culture model of bone marrow mesenchymal stem cell sheet and umbilical vein endothelial cells: establishment and identification

Zhou Zhen, Wang Yamin, Ren Fei, Zhang Zhaoqiang, Zeng Shuguang, Yang Xi
  

  1. Stomatology Hospital, Southern Medical University, Guangzhou 510280, Guangdong Province, China
  • Revised:2019-04-02 Online:2019-09-08 Published:2019-09-08
  • Contact: Wang Yamin, Master, Attending physician, Stomatology Hospital, Southern Medical University, Guangzhou 510280, Guangdong Province, China; Ren Fei, Chief physician, Stomatology Hospital, Southern Medical University, Guangzhou 510280, Guangdong Province, China
  • About author:Zhou Zhen, Master, Attending physician, Stomatology Hospital, Southern Medical University, Guangzhou 510280, Guangdong Province, China
  • Supported by:

    the Self-Financing Science and Technology Research Project of Guangdong Province, No. 2017ZC0124 (to ZZ); Guangdong Medical Science and Technology Research Project, No. C2017061 (to RF); the Natural Science Foundation of Guangdong Province, No. 2018A030313759 (to ZZQ); Guangzhou Science and Technology Plan Project, No. 201707010193 (to ZSG)

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摘要：

文章快速阅读：

文题释义：
细胞膜片技术：是通过物理的方法将扩增融合的细胞从培养皿底壁分离，获得膜片状细胞结构的一种技术方法。一方面不用对细胞进行胰蛋白酶消化，既保持了细胞的活性又提高了细胞接种浓度，含有丰富的细胞外基质；另一方面保留了体外培养过程中细胞自身分泌的细胞外基质，细胞自身作为种子细胞，更接近人的正常组织。
间充质干细胞：属于中胚层的一类多能干细胞，具有来源广泛、强大的增殖能力、多向分化潜能及免疫排斥反应低等特性，作为种子细胞构建细胞膜片具有广阔的应用前景。

 

摘要
背景：细胞膜片技术是近年来热门的组织工程学技术，但是膜片中间部位的营养供给和移植后血供问题易导致失败，所以预血管化是提高组织工程材料移植后存活率的关键。
目的：比较在不同氧含量下，人骨髓间充质干细胞和脐静脉内皮细胞共培养形成预血管化骨髓间充质干细胞膜片的生物学性能。
方法：实验分2组：常氧共培养组即常氧(体积分数20%O2)条件下形成人骨髓间充质干细胞膜片，常氧条件下加入人脐静脉内皮细胞共培养；低氧共培养组，即低氧(体积分数2%O2)条件下形成人骨髓间充质干细胞膜片，常氧条件下加入人脐静脉内皮细胞共培养。采用免疫荧光染色技术观察并比较共培养7 d两组的微血管生成情况，并通过ELISA法比较不同氧含量下骨髓间充质干细胞膜片内血管内皮生长因子的水平。
结果与结论：①与常氧共培养组比较，低氧共培养组的血管更长、血管密度更高、管间交互连接更多(P < 0.05)；②低氧共培养组的血管内皮生长因子水平高于常氧共培养组(P < 0.05)；③结果证实，低氧共培养组较常氧共培养组膜片更利于血管生长，对于建立优化的预血管化膜片具有可行性。

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
ORCID: 0000-0002-0987-1211(周震)

关键词: 低氧, 常氧, 细胞膜片, 骨髓间充质干细胞, 免疫荧光染色, 脐静脉内皮细胞, 血管内皮生长因子, 干细胞, 血管化, 组织工程

Abstract:

BACKGROUND: Cell sheet technology is a recent popular technique for tissue engineering. However, a failure is prone to occur because of poor nutrient supply in the middle part of the membrane and post-transplantation blood supply, so pre-vascularization determines the survival rate of tissue engineering materials after transplantation.
OBJECTIVE: To investigate the pre-vascularization of bone marrow mesenchymal stem cell sheets co-cultured with human umbilical vein endothelial cells under different oxygen tensions.
METHODS: Bone marrow mesenchymal stem cell sheets were prepared under physiological hypoxia (2% O2) and normoxia (20% O2), and then co-cultured with human umbilical vein endothelial cells under normoxia. At 7 days of co-culture the angiogenesis was observed and compared by immunofluorescence staining. The level of vascular endothelial growth factor was compared under different oxygen tensions using ELISA.
RESULTS AND CONCLUSION: Compared with the normoxia co-culture group, there were longer microvessels, higher vessel density and more microvessel networks in the hypoxia co-culture group (P < 0.05). The expression of vascular endothelial growth factor in the hypoxia co-culture group was higher than that in the normoxia co-culture group (P < 0.05). Therefore, the hypoxia co-culture is more beneficial to the growth of microvessels than the normoxia co-culture, and it is feasible to construct an optimized prevascularized cell sheets.

Key words: hypoxia, normoxia, cell sheet, bone marrow mesenchymal stem cells, immunofluorescence staining, umbilical vein endothelial cells, vascular endothelial growth factor, stem cells, vascularization, tissue engineering

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