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Most murine embryonic stem cell lines have been derived from the inner cell mass of blastocysts and extensively studied in different aspects including generation of organ specific cells. However, no detailed studies have been made on cardiac specific gene expression, immunocytochemical and electrophysiological characterisation of cardiomyocytes generated from early stage (preimplantation) embryo derived embryonic stem cells in mice. In the present study, new embryonic stem cell lines were derived from early stage preimplanatation embryos in mice. In vitro differentiation of such cell lines readily generated cardiomyocytes, which expressed different cardiac specific genes in a temporally regulated manner as well as cardiac cells specific proteins. This is probably the first report, which showed the temporal pattern of cardiac specific genes as well as protein expression in cardiac cells generated from in vitro differentiation of preimplantation embryo derived ES cells