关键词: 心脏成纤维细胞, 原代培养, 裸鼹鼠

Abstract: Objective To establish the method for the isolation, culture and identification of cardiac fibroblasts from naked mole rat suckling mice in vitro. Method Neonatal rat hearts were isolated on ice from naked mole rats born 1 to 3 days after birth. The hearts were cut into 1 mm×1 mm×1 mm pieces with ophthalmic scissors. Cardiac fibroblasts were isolated by digestion with trypsin and type II collagenase successively. Morphological characteristics of cardiac fibroblasts were observed under light microscope, proliferation activity was detected by CCK-8 assay, vimentin expression was detected by Western blot and immunofluorescence assay. Result Cardiac fibroblasts isolated by enzyme digestion were completely adherent in primary culture for 4 h, and the whole 35 mm dish could be covered in 36 h. The proliferation activity of cardiac fibroblasts was the highest after 2 to 3 days of subculture, and the expression of vimentin was confirmed by Western blot and immunofluorescence. Conclusion Cardiac fibroblasts isolated by trypsin and type II collagenase digestion can obtain cardiac fibroblasts with good viability and high purity.

Key words: cardiac fibroblasts, primary culture, naked mole rats