TY - JOUR
T1 - Influence of Electromechanical Activity on Cardiac Differentiation of Mouse Embryonic Stem Cells
AU - Limpitikul, Worawan
AU - Christoforou, Nicolas
AU - Thompson, Susan A.
AU - Gearhart, John D.
AU - Tung, Leslie
AU - Lipke, Elizabeth A.
PY - 2010/9
Y1 - 2010/9
N2 - opment. Electrical pacing was applied to confluent monolayers of mESC-CMs during late-stage differentiation (days 16-18). Alternatively, spontaneous contraction was suppressed by (a) blocking ion currents with CsCl (HCN channel), trazodone (T-type Ca2+ channel), or both CsCl and trazodone on days 11-18; or (b) applying blebbistatin (excitation-contraction uncoupler) on days 11-14. Electrophysiological properties and gene expression were examined on day 19 and 18, respectively. Optical mapping revealed no significant difference in conduction velocity (CV) in paced vs. non-paced monolayers, nor were there significant changes in gene expression of connexin-43, Na-Ca exchanger (NCX), or myosin heavy chain (MHC). However, CV variability among differentiation batches and CV heterogeneity within individual monolayers were significantly lower in paced mESC-CMs. Alternatively, while the four drug treatments suppressed contraction with varying degrees (up to complete inhibition), there was no significant difference in CV for any of the treatments compared with controls. Trazodone treatment significantly reduced CV variability as compared to controls, whereas CsCl treatment significantly reduced CV heterogeneity. Distinct changes in gene expression of connexin-43, MHC, HCNl, Cav3. 1/3.2 were not observed. Electrical pacing, but not suppression of spontaneous contraction, during late-stage differentiation reduces the intrinsic variability of CV among differentiation batches and across individual monolayers, which can be beneficial in the application of ESCs for myocardial tissue repair.
AB - opment. Electrical pacing was applied to confluent monolayers of mESC-CMs during late-stage differentiation (days 16-18). Alternatively, spontaneous contraction was suppressed by (a) blocking ion currents with CsCl (HCN channel), trazodone (T-type Ca2+ channel), or both CsCl and trazodone on days 11-18; or (b) applying blebbistatin (excitation-contraction uncoupler) on days 11-14. Electrophysiological properties and gene expression were examined on day 19 and 18, respectively. Optical mapping revealed no significant difference in conduction velocity (CV) in paced vs. non-paced monolayers, nor were there significant changes in gene expression of connexin-43, Na-Ca exchanger (NCX), or myosin heavy chain (MHC). However, CV variability among differentiation batches and CV heterogeneity within individual monolayers were significantly lower in paced mESC-CMs. Alternatively, while the four drug treatments suppressed contraction with varying degrees (up to complete inhibition), there was no significant difference in CV for any of the treatments compared with controls. Trazodone treatment significantly reduced CV variability as compared to controls, whereas CsCl treatment significantly reduced CV heterogeneity. Distinct changes in gene expression of connexin-43, MHC, HCNl, Cav3. 1/3.2 were not observed. Electrical pacing, but not suppression of spontaneous contraction, during late-stage differentiation reduces the intrinsic variability of CV among differentiation batches and across individual monolayers, which can be beneficial in the application of ESCs for myocardial tissue repair.
KW - Cardiac regeneration
KW - Cell culture
KW - Electrical stimulation
KW - Electrophysiology
KW - Optical mapping
UR - http://www.scopus.com/inward/record.url?scp=79953184629&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=79953184629&partnerID=8YFLogxK
U2 - 10.1007/s13239-010-0020-8
DO - 10.1007/s13239-010-0020-8
M3 - Article
C2 - 29057018
AN - SCOPUS:79953184629
SN - 1869-408X
VL - 1
SP - 179
EP - 193
JO - Cardiovascular Engineering and Technology
JF - Cardiovascular Engineering and Technology
IS - 3
ER -