Comparing the Rate of Proliferation and Cardiac Differentiation between B6- and MRL- Mouse Embryonic Stem Cells Previous

Previous studies crowned the MRL mouse as a â€oesuper healing― strain due to its superior capacities to fully regrow an ear punch wound and repair damaged bone cartilage; however, the ability to regenerate cardiac tissue following myocardial infarction remains to debate. The present study aims to determine whether mouse embryonic stem cells (mESCs) derived from the MRL mouse strain (MRL-mESCs) will have a higher proliferation rate than those derived from the most commonly used B6 mouse strain (B6-mESCs). Additionally, we will determine whether MRL-mESCs have a higher efficiency of cardiac differentiation into beating cardiomyocytes (CMs). To this end, both MRL- and B6-mESC lines were cultured on gelatin-coated plates with daily mESC medium change. To remove any potential contamination of fibroblasts originally from the mouse embryonic feeder layer, the mESC colonies were mechanically dissected with a needle under a microscope. These pure colonies were passaged every 3-5 days, at which time they were counted to determine proliferation rate. To induce cardiac differentiation, mESCs were seeded in a 96-well conical tube array and cultured for 5 days to form embryoid bodies (EBs). The EBs were then plated and driven toward the cardiac lineage with cardiac differentiation medium. 14 days after the EBs were plated, they were examined to determine the efficiency of cardiac differentiation. Our preliminary data indicates that B6-mESCs appear to have a higher proliferation rate than the MRL cells. The ongoing experiments aim to increase the sample size for statistical analysis