Chemical induction of cardiac differentiation in p19 embryonal carcinoma stem cells.
P19 cells, a pluripotent cell line derived from a teratocarcinoma induced in C3H/HeHa mice, have been widely used as a model system to study cardiac differentiation. We have used these cells to evaluate the extent to which exposure to DMSO and/or cardiogenol C for 4 days in suspension culture enhanced their differentiation into cardiomyocytes. Cardiac differentiation was assessed by observing beating clusters and further confirmed using immunocytochemical, biochemical, and pharmacological approaches. The presence of functional gap junctions in differentiated P19 cells was identified through calcium wave analyses. Proliferation rate and cell death were analyzed by BrdU incorporation and activated caspase-3 immunodetection, respectively. Beating clusters of differentiated P19 cells were only found in cultures treated with DMSO. In addition, groups treated with DMSO up-regulated cardiac troponin-T expression. However, when DMSO was used together with cardiogenol C the up-regulation was less than that with DMSO alone, approximately 1.5 times. Moreover, P19 cells cultured in DMSO or DMSO plus 0.25 microM cardiogenol C had lower proliferation rates and higher numbers of activated caspase-3-positive cells. In summary, using several methodological approaches we have demonstrated that DMSO can induce cardiac differentiation of P19 cells but that cardiogenol C does not.
Jasmin
Instituto de Biofísica Carlos Chagas Filho, UFRJ, Rio de Janeiro, Brazil.
Spray
David C
DC
Campos de Carvalho
Antonio Carlos
AC
Mendez-Otero
Rosalia
R
D000814:Aniline Compounds / Q000494:pharmacology*; D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D015153:Blotting, Western; D020013:Calcium Signaling / Q000187:drug effects; D053148:Caspase 3 / Q000378:metabolism; D002454:Cell Differentiation / Q000187:drug effects*; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D004121:Dimethyl Sulfoxide / Q000494:pharmacology*; D004357:Drug Synergism; D054278:Embryonal Carcinoma Stem Cells / Q000378:metabolism / Q000473:pathology; D004789:Enzyme Activation / Q000187:drug effects; D051379:Mice; D008809:Mice, Inbred C3H; D018613:Microscopy, Confocal; D032383:Myocytes, Cardiac / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D050814:Octamer Transcription Factor-3 / Q000096:biosynthesis; D011743:Pyrimidines / Q000494:pharmacology*; D020107:Troponin T / Q000096:biosynthesis; D015854:Up-Regulation / Q000187:drug effects
D000814:Aniline Compounds; D050814:Octamer Transcription Factor-3; C494174:Pou5f1 protein, mouse; D011743:Pyrimidines; D020107:Troponin T; C484397:cardiogenol C; D053148:Caspase 3; D004121:Dimethyl Sulfoxide