大鼠肺微血管内皮细胞的体外分离培养与纯化

doi:10.11843/j.issn.0366-6964.2016.10.024

Abstract

Abstract:

This study aims at establishing a simple method of obtaining the high-purity rat pulmonary microvascular endothelial cells isolated and cultured in vitro.Peripheral lung tissues were separated from 5 to 7 days of SD rat under sterile condition.Rat pulmonary microvascular endothelial cells was acquired by taking the tissue graft block method，digested with trypsin and then maintained at 37 ℃ in a 5% CO₂ humidified atmosphere.The cells were purificated using the method of immune magnetic beads，subsequently identified by the immunofluorescence and flow cytometry，and then determined by MTT colorimetric analysis to detect the original generation of pulmonary endothelial cell growth curve.Rat lung microvascular endothelial cells was successfully obtained by taking the tissue graft block method and individual cells under inverted microscope showed short fusiform or polygon and a typical paying stone appearance with single-layer adherent growth after confluence.The cells were strongly positive for CD31 factor and negative（P=0.1，and P<0.5） for lymphatic endothelial cell specific surface marker VEGFR-3 detected by immunofluorescence staining and flow cytometry respectively.In our study，a method of higher purity of rat lung microvascular endothelial cells culture in vitro was successfully established.

Key words: rats, pulmonary microvascular endothelial cells, isolation and cultivation in vitro, immune magnetic beads

CLC Number:

S852.21

LIANG Hong-wei，FENG Bo，ZHU Wen-yu，WANG Jian-fang，ZHANG Tao，MU Xiang. Cultivation and Purification of Rat Pulmonary Microvascular Endothelial Cells in vitro[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2016, 47(10): 2143-2150.

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Cultivation and Purification of Rat Pulmonary Microvascular Endothelial Cells in vitro

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