Primary culture and identification of embryonic rat cardiac fibroblasts
SHEN Jun
YUAN Ping
TANG Jun-ming
YANG Jian-ye
LI Xing-yuan
ZHANG Lei
ZHAO Ji-xian
ZHANG Huan-xin
XUE Shi-zhen
FENG Yi
WANG Jia-ning
Abstract：Objective To establish a suitable method for the isolation, culture and identification of embryonic rat cardiac fibroblasts in vitro.Methods Embryos of pregnant SD rats (19 days) were taken to get its heart, then the heart were cut into meat paste and digested with trypsinase plus collagenase Ⅱ/Ⅳ to isolate cardiac fibroblasts. Cardiac fibroblasts from embryonic rat hearts were harvested by differential adherent separation. The morphological changes of cardiac fibroblasts were observed under light microscope. The second generation of cardiac fibroblasts was identified by immunofluorescence with vimentin, desmin, von Willebrand factor (vWF) and smooth muscle actin (α-SMA).Results The primary culture of cardiac fibroblasts isolated by differential adherence method was completely adherent for 120 min, and the cells were cultured well in the third to fifth passage. The purity of cardiac fibroblast vimentin immunofluorescence was up to 97% after 24 hours. Cultured for 2 to 3 days, the proliferation of cardiac fibroblasts was the strongest. Conclusion Differential adherence assay combined with immunofluorescence assay can be used to separate embryonic rat cardiac fibroblasts. A great quantity, purity and high survival rate of cardiac fibroblasts can be effectively cultured by this method. And it is useful to explore the pathologic mechanisms of cardiovascular disease in cellular level.
Keywords：Cardiac fibroblastsPrimary cultureEmbryonic ratIdentification
Publication Date：2016-01-01
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：4( 1441-1444 )