Enrichment of cardiomyocytes derived from mouse embryonic stem cells.
BACKGROUND
Embryonic stem (ES) cell-derived cardiomyocytes transplantation and tissue engineering together represent a promising approach for the treatment of myocardial infarction, despite the limited supply of cardiac myocytes. This study examines whether functional cardiomyocytes can be efficiently enriched from mouse embryonic stem (mES) cells.


METHODS
mES cells were induced by ascorbic acid to differentiate into cardiomyocytes. Beating cells were observed after 1 week and increased in number with time while under differentiation conditions. Furthermore, the differentiated cultures could be dissociated and enriched by Percoll gradient density centrifugation.


RESULTS
The beating cells expressed markers characteristic of cardiomyocytes, such as cardiac troponin T (cTnT). The enriched population contained 88.7% cardiomyocytes and showed expression of cardiomyocyte markers of troponin T and cardiac genes, including alpha-MHC, beta-MHC, ANF and Nkx2.5. However, Oct-4, a marker of early-stage ES cells, was not expressed in the mES cell-derived cardiac cell clusters. Moreover, the mES cell-derived and Percoll-enriched cardiomyocytes responded appropriately to cardioactive drugs, as did normal neonatal rat cardiomyocytes.


CONCLUSIONS
mES cell-derived functional cardiomyocytes can be enriched by the method of discontinuous Percoll gradient centrifugation. The ability to differentiate and enrich for functional mouse cardiomyocytes makes it possible for further development of these cells as a model of myocardial repair through cell transplantation or tissue engineering.
E
Ling-Ling
LL
Department of Tissue Engineering Center, Beijing Institute of Basic Medical Sciences and Tissue Engineering Research Center, Academy of Military Medical Sciences, Beijing, PR China.
Zhao
Yun-Shan
YS
Guo
Xi-Min
XM
Wang
Chang-Yong
CY
Jiang
Hong
H
Li
Jing
J
Duan
Cui-Mi
CM
Song
Ying
Y
D000818:Animals; D000831:Animals, Newborn; D001205:Ascorbic Acid / Q000494:pharmacology; D019149:Bioreactors; D002121:Calcium Channel Blockers / Q000494:pharmacology; D002454:Cell Differentiation / Q000187:drug effects / Q000502:physiology; D002478:Cells, Cultured; D002499:Centrifugation, Density Gradient; D003102:Colloids / Q000494:pharmacology; D004110:Diltiazem / Q000494:pharmacology; D018507:Gene Expression Regulation, Developmental; D007150:Immunohistochemistry; D051379:Mice; D018613:Microscopy, Confocal; D032383:Myocytes, Cardiac; D050814:Octamer Transcription Factor-3 / Q000378:metabolism; D011205:Povidone / Q000494:pharmacology; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012822:Silicon Dioxide / Q000494:pharmacology; D013234:Stem Cells / Q000166:cytology* / Q000187:drug effects; D023822:Tissue Engineering / Q000379:methods*
D002121:Calcium Channel Blockers; D003102:Colloids; D050814:Octamer Transcription Factor-3; C016039:Percoll; D012822:Silicon Dioxide; D004110:Diltiazem; D011205:Povidone; D001205:Ascorbic Acid