Simultaneous isolation of myocardial cells and cardiac fibroblasts from neonatal rats

doi:10.3969/j.issn.2095-4344.2013.24.007

Abstract

Abstract:

BACKGROUND: Myocardial cells and cardiac fibroblasts are simultaneously isolated through optimizing the culture conditions of myocardial cells, by use of the variations of adhesion properties of myocardial cells.
OBJECTIVE: To explore the co-separation method for myocardial cells and cardiac fibroblasts in newborn rats.
METHODS: Myocardial tissue in newborn rats was digested with low concentration trypsin at cold conditions and with collagenase II at 37 ℃. The myocardial cells and cardiac fibroblasts were isolated with differential centrifugation method, and myocardial cell culture conditions were improved using different kinds of serum. Immunofluorescence staining was used to evaluate the purity of myocardial cells and cardiac fibroblasts.
RESULTS AND CONCLUSION: The myocardial Troponin T positive cell rate was 89.3% at culture 48 hours, and myocardial fibroblast vimentin positive cell rate was 93.6% at passage 2. The percentage of positive myocardial cells cultured with medium containing horse serum was obviously higher than that with bovine serum culture (P < 0.05). This isolation method of rat myocardial cells can gain myocardial cells and cardiac fibroblasts at the same time, with high purity and good activity.

Key words: tissue construction, heart tissue construction, myocardial cells, cardiac fibroblasts, trypsin, collagenase, combined digestion, cell culture, horse serum, bovine serum, proliferation, differentiation

CLC Number:

R318

Shi Yu-lu, Li Xiao-yuan, Cao Mei-na, Yu Shu-yuan, Wang Ping. Simultaneous isolation of myocardial cells and cardiac fibroblasts from neonatal rats[J]. Chinese Journal of Tissue Engineering Research, 2013, 17(24): 4414-4420.

Figures/Tables 4

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