基础医学与临床 ›› 2022, Vol. 42 ›› Issue (5): 714-720.doi: 10.16352/j.issn.1001-6325.2022.05.027

• 研究论文 • 上一篇    下一篇

血脑屏障体外细胞模型的建立与比较

苏钰雯, 修建波, 许琪*   

  1. 中国医学科学院基础医学研究所 北京协和医学院基础学院 生物化学与分子生物学系医学分子生物学国家重点实验室,北京 100005
  • 收稿日期:2022-02-07 修回日期:2022-03-22 出版日期:2022-05-05 发布日期:2022-04-28
  • 通讯作者: * xuqi@pumc.edu.cn
  • 基金资助:
    国家自然科学基金(81930104,31970952);国家重点研发计划(2020YFA0804502);中国医学科学院创新工程项目 (2021-1-I2M-020)

Establishment and comparison of in vitro blood-brain barrier cell models

SU Yu-wen, XIU Jian-bo, XU Qi*   

  1. State Key Laboratory of Medical Molecular Biology, Department of Molecular Biology and Biochemistry, Institute of Basic Medical Sciences CAMS, School of Basic Medicine PUMC, Beijing 100005, China
  • Received:2022-02-07 Revised:2022-03-22 Online:2022-05-05 Published:2022-04-28
  • Contact: * xuqi@pumc.edu.cn

摘要: 目的 构建不同培养模式的血脑屏障(BBB)体外细胞模型并进行结构和功能的比较和分析。方法 用人脑微血管内皮细胞系(hCMEC/D3)、人脑星形胶质母细胞瘤细胞系(U87MG)和人脑血管周细胞系(HBVP)3种细胞在Transwell装置中构建细胞模型。根据细胞种类和细胞位置的不同,将模型分为单培养、双培养(hCMEC/D3+ U87MG或HBVP)、不接触共培养、半接触共培养和全接触共培养5种血脑屏障体外细胞模型。检测模型的通透性和渗透性以及相关蛋白和基因的表达,比较不同模型的结构和功能特点并分析。结果 与单培养和双培养模型相比,共培养模型的渗透性降低,紧密性升高,血脑屏障相关的基因和蛋白的表达也升高(P<0.05)。在共培养模型中,半接触和全接触培养模型具有更低的渗透性(P<0.001),全接触细胞培养模型的紧密性更高(P<0.01),部分血脑屏障相关蛋白和基因的表达也更高(P<0.05)。结论 全接触共培养细胞模型具有更优的血脑屏障相关性质,更适用于血脑屏障研究。

关键词: 内皮细胞, 星形胶质细胞, 周细胞, 血脑屏障, 共培养

Abstract: Objective To construct and evaluate the in vitro models of blood-brain barrier (BBB)and to analyze their structural and functional characteristics. Methods Human microvascular endothelial cell line(hCMEC/D3), Uppsala 87 malignant glioma cell line(U87MG) and human brain vascular pericyte(HBVP) cell line were used to construct blood-brain barrier cell models. The models were divided into single-culture model, bi-culture (hCMEC/D3+U87MG or HBVP), no-contact co-culture model, semi-contact co-culture and full-contact co-culture model according to different cell types and cell locations. The permeability and tightness of the models as well as the expression of related proteins and genes were detected for comparing and analyzing the structural and functional characteristics of different models. Results Compared with the single-culture model and the bi-culture model, the permeability and compactness of the co-culture model decreased and the expression of BBB-related genes and proteins also increased(P<0.05). In the co-culture models, full-contact co-culture model showed higher compactness(P<0.01) and higher BBB-related protein and gene expression. Semi-contact and full-contact culture model had lower permeability(P<0.05). Conclusions Full-contact co-culture model has better BBB-related properties and is more suitable for BBB research.

Key words: endothelial cell, astrocyte, pericyte, blood-brain barrier, co-culture

中图分类号: