Previously, our team was limited to performing RNA-seq on nuclei derived from ACMs. To overcome this restriction, we used Parse Biosciences’ Evercode assay on whole cardiomyocyte cells. A high number of genes and transcripts per cell were identified. Beginning with a protocol our lab developed for the isolation of ventricular cells, we found strong expression of canonical markers for ventricular ACMs (Fig. 2) and are excited to use Evercode in our future studies.
The heart was harvested from a nine-month-old male mouse (wildtype, C57BL/6). Ventricular cardiomyocytes were isolated using the Langendorff perfusion digestion procedure. The cell suspensions were then passed through a 100 μm cell strainer to remove tissue debris and purified by low-speed centrifugation, resulting in approximately 95% pure adult cardiomyocytes (ACMs) (Fig.1, scale bar = 100 um).
Figure 1, Adult cardiomyocytes (ACMs)
Figure 2, Canonical markers for ventricular ACMs