Myosin heavy chain expression in contracting myocytes isolated during embryonic stem cell cardiogenesis.
Mouse embryonic stem (ES) cells are totipotent cells derived from the inner cell mass of the preimplantation blastocyst and are capable of differentiating in vitro into cardiac myocytes. Attached cultures of differentiating ES cells were established to document the timing of contractile development by microscopic observation and to permit the microdissection of cardiac myocytes from culture. The onset of spontaneous contraction varied markedly in differentiation culture, with contraction being maintained on average for 9 days (range, 1 to 75 days). Indirect immunofluorescence in microscopy showed that myosin expression was localized to the contracting cardiac myocytes in culture. Myosin heavy chain (MHC) isoform expression in microdissected ES cell-derived cardiac myocytes was determined by means of sodium dodecyl sulfate-polyacryl-amide gel electrophoresis. The distribution of MHC isoform expression in isolated ES cell cardiac myocytes was as follows: 27% expressed the beta-MHC isoform, 33% expressed both the alpha- and beta-MHC isoforms, and 40% expressed the alpha-MHC isoform. MHC phenotype was correlated to the duration of continuous contractile activity of the myocytes. Myocytes that had just initiated spontaneous contractile activity predominantly expressed the beta-MHC (average days of contraction before isolation, 2.5 +/- 0.7). The alpha-MHC isoform was detected after mouse prolonged contractile activity in vitro (1 to 5 weeks). A strong correlation was obtained between MHC phenotype and days of contraction of the cardiac myocyte preparations isolated from ES cell cultures (r = .93). The apparent transition in MHC isoform expression during ES cell differentiation parallels the beta- to alpha-MHC isoform transition characteristic of murine cardiac development in vivo. These findings are evidence that ES cell cardiac myocyte differentiation follows the normal developmental program of murine cardiogenesis.
Metzger
J M
JM
Department of Physiology, School of Medicine, University of Michigan, Ann Arbor 48109-0622, USA.
Lin
W I
WI
Johnston
R A
RA
Westfall
M V
MV
Samuelson
L C
LC
D000818:Animals; D002454:Cell Differentiation; D002478:Cells, Cultured; D006321:Heart / Q000196:embryology* / Q000502:physiology; D051379:Mice; D009200:Myocardial Contraction; D009218:Myosins / Q000096:biosynthesis*; D013234:Stem Cells / Q000378:metabolism
D009218:Myosins