When can I assay or perform gene expression analysis on your cardiomyocytes?

iCell® Cardiomyocytes remain very stable over time in culture as demonstrated by Babiarez et al. 2012, but the cryopreservation and thawing process can induce a temporary stress response. Reanimation of iCell Cardiomyocytes generally takes several days for them to return to a steady state, which requires time for the cells to attach, spread out, and form a synchronous beating monolayer.

While assay timing is assay dependent, our data generally indicate that iCell Cardiomyocytes can be used for biochemical and transfection assays after Day 7 post-plating and electrophysiological or contraction assays after 10 - 14 days post-plating. For electrophysiology and contraction assays, it is critical to wait for appropriate gap junctions to form between cells to establish a syncytium and observe uniform beating.

Gene expression in iCell Cardiomyocytes will be incredibly stable over time and mimic what you see for human adult cardiac tissue as examined by Puppala et al. 2013. However, we have observed some changes in gene expression associated with a stress response shortly after thawing the cells that peaks in the first few days and then recedes after several days as the cells return to their steady state.

iCell Cardiomyocytes have been cultured for >120 days (up to 9 months). For more details about how long cardiomyocytes can be maintained in culture, see Solution 00000019B.

Reference:

  1. iCell Cardiomyocytes User's Guide
  2. Babiarz et al. 2012. Determination of the human cardiomyocyte mRNA and miRNA differentiation network by fine-scale profiling
  3. Puppala et al. 2013. Comparative gene expression profiling in human-induced pluripotent stem cell–derived cardiocytes and human and cynomolgus heart tissue