Original Pre-Clinical ScienceCoculturing with endothelial cells promotes in vitro maturation and electrical coupling of human embryonic stem cell–derived cardiomyocytes
Section snippets
Maintenance of hESC lines
The hESC cell lines with a green fluorescent protein (GFP) reporter for the cardiac gene, Nkx2.5, were obtained as previously described (NKX2-5GFP hESC).9 The hESC cell lines ES4 were purchased from Wicell Research Institute. The permissions for use of these cell lines were obtained after review by the Cornell-Rockefeller-Sloan Kettering Institute ESC Research Oversight Committee.
The hESCs were grown on feeder layer free conditions on growth factor-reduced BD Matrigel (#354230, BD Biosciences)
Cardiac differentiation optimization
We first used classical 2D cultures to derive cardiomyocytes from hESCs with or without an ECs feeder layer. The presence of endothelium induced increased differentiation (13.5% ± 1.5% in the control vs 18.6% ± 1.2%) with the ECs, as defined by the number of differentiated hESCs expressing NKX2-5GFP over the total number of hESCs as determined by flow cytometry (p < 0.01; Supplementary Figure 1A–C, available online at www.jhltonline.org); however, the efficiency of this protocol was quite low,
Discussion
Here, we demonstrated that the in vitro use of an activated endothelial feeder increased the differentiation of cardiomyocytes from hESCs and resulted in a differentiated cardiomyocyte population with a synchronous beating pattern that may be more adapted to clinical use.32, 33
Disclosure statement
None of the authors has a financial relationship with a commercial entity that has an interest in the subject of the presented manuscript or other conflicts of interest to disclose.
The authors thank the Flow Cytometry Facility within the Microscopy Core at Weill Cornell Medical College in Qatar for contributing to these studies. The Core is supported by the “Biomedical Research Program at Weill Cornell Medical College in Qatar," a program funded by Qatar Foundation. The authors acknowledge
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These authors contributed equally to this work.