An improved protocol for primary culture of cardiomyocyte from neonatal rat / 中华心血管病杂志

ABSTRACT

<p><b>OBJECTIVE</b>The present study describes an improved in vitro culture method for obtaining high purity, vital and fully functional cardiomyocytes from neonatal rat.</p><p><b>METHODS</b>After cutting ventricular tissue with improved method, ventricular tissues were digested with low concentrations of trypsin overnight at 4 °C, and then underwent collagenase II digestion. Thereafter, cardiomyocytes were purified by combined differential adhesion and chemical inhibition methods.</p><p><b>RESULTS</b>Adherent cardiomyocytes were seen at 12 h after culture, spontaneously beating cardiomyocytes were observed at 24 h after culture, crosslinked cardiomyocytes were found at 48 h after culture, adhesion clustered cardiomyocytes were seen at 72 h after culture, dense network formed from inter-connected was evidenced together with radial arranged cell clusters and cell pseudopodia 96 h the mutual contact woven into and formed radically ordering cell clusters and island-like beating cardiomyocytes at 96 h after culture. The cell survival rate and purity were more than 98%.</p><p><b>CONCLUSION</b>Fully functional spontaneous beating cardiomyocytes can be obtained by the use of this improved primary neonatal rat cardiomyocytes culture method.</p>