Can I use iCell Cardiomyocytes2 to model cardiac hypertrophy?
Yes, iCell® Cardiomyocytes2 recapitulate numerous properties of native cardiac myocytes, including a hypertrophic response. This makes them an excellent in vitro model of cardiac hypertrophy for multiple endpoints as demonstrated reproducibly in a peer-reviewed publication by Carlson et al. (2013) and in optimized application protocols using endothelin-1 to induce a hypertrophic response, measured by flow cytometry with NPPB/BNP.
General Protocol Recommendations:
- Assay timing: Culture iCell Cardiomyocytes2 according to the appropriate Hypertrophy Application Protocol (see links below). Induce with endothelin-1 (ET-1) on day 5 post-thaw and evaluate on day 6 post-thaw following an overnight induction (~18 hours). Note that this workflow recommendation starts on a Wednesday (day 0), avoids culturing the cells over the weekend, and requires ET-1 induction on a Monday (day 5) and assaying on Tuesday (Day 6).
- Assay density: 50,000 viable cells/well in 96-well plate.
- Assay ECM: 5 µg/ml fibronectin (100 µl/well, coat for 1 hour at 37°C)
- Assay media:
- Plating Medium: iCell Cardiomyocytes Plating Medium (at thaw)
- Maintenance Medium: iCell Cardiomyocytes Maintenance Medium (at 4 hours post-thaw), and then perform a 100% medium exchange at 48 hours post-thaw) to serum-free Supplemented William's E Medium (SWE, supplemented with a 1:25 dilution of Cocktail B)
Application Protocols for modeling cardiac hypertrophy with ET-1 induction:
- Application Protocol: Modeling Cardiac Hypertrophy: Endothelin-1 Induction with ELISA Analysis Application Protocol
- Application Protocol: Modeling Cardiac Hypertrophy: Endothelin-1 Induction with Flow Cytometry Analysis
- Application Protocol: Modeling Cardiac Hypertrophy: Endothelin-1 Induction with High Content Analysis
- Application Protocol: Modeling Cardiac Hypertrophy: Endothelin-1 Induction with qRT-PCR Analysis
- Carlson et al. (2013) Phenotypic Screening with Human iPS Cell-derived Cardiomyocytes: HTS-compatible Assays for Interrogating Cardiac Hypertrophy