血管内皮细胞和心脏组织块立体培养.pdfVIP

生理学报 Acta Physiologica Sinica, April 25, 2005, 57 (2): 259-269 259 实验技术 血管内皮细胞和心脏组织块的立体培养 王铭洁，蔡文杰，姚　泰，朱依纯* 复旦大学上海医学院生理学与病理生理学系，上海 200032 摘 要：本文旨在对比研究二维平面与三维立体培养模式下，内皮细胞和心脏组织形态学的差异。采用胶内、胶上、三 明治模式、玻片培养小室模型等多种Ⅰ型胶原立体培养模型，通过免疫荧光技术及显微形态学观察组织和细胞的生长情况。 在二维平面培养中，原代心脏血管内皮细胞呈铺路石样排列；而在三维胶原培养模式中，内皮细胞呈长梭状形态，并迁 入胶原培养介质中，和体内血管新生及血管生成过程中的内皮细胞活化表型相似。加入血管内皮生长因子(vascular endo- thelial growth factor, VEGF) 能增强内皮细胞管状结构的形成。在三维胶原中，心脏组织块生长良好，迁出的细胞将相邻 组织块连接起来，组织块有自发的搏动。本工作表明，改进的薄层胶原培养、玻片培养小室模型和动脉条模型是较好的 研究血管生成和血管新生的工具。在三维培养的情况下，内皮细胞通过空间增殖、迁移和锚定，可形成管状结构，比 二维平面培养更适合用于血管新生的研究。不同的立体培养模型可用于不同目的的研究。 关键词：血管新生；Ⅰ型胶原；内皮细胞；组织培养 中图分类号：Q463; R33 Endothelial cells and cardiac explants in three-dimensional culture system WANG Ming-Jie, CAI Wen-Jie, YAO Tai, ZHU Yi-Chun* Department of Physiology and Pathophysiology, Shanghai Medical College, Fudan University, Shanghai 200032, China Abstract: To observe the morphological features of endothelial cells and cardiac explants cultured in two- or three-dimensional culture systems, several three-dimensional collagen type Ⅰ culture systems, such as the in gel, on gel, sandwich model, and the microscope slide model, were used to examine the growth patterns of the cells and explants from heart by using immunofluorescence staining and microscopic observation in the presence or absence of vascular endothelial growth factor (VEGF). In two-dimensional cultures the primary cardiac endothelial cells arrayed into a cobblestone-like structure. When cultured in three-dimensional matrix, the cells were elongated and migrated into the gel, with a phenotype similar to that in the process of angiogenesis and vasculogenesis in vivo. VEGF promoted the process of the endothelial cells transforming into tube-like structure. Cardiac explants grew well in the collagen gel. Adjacent explants were connected to each other by the migrating cells with the occurrence of autorhythmic beating of the explan