© 1995 American Heart Association, Inc.
Articles |
Myosin Heavy Chain Expression in Contracting Myocytes Isolated During Embryonic Stem Cell Cardiogenesis
From the Department of Physiology, School of Medicine, University of Michigan, Ann Arbor.
Correspondence to Joseph M. Metzger, Department of Physiology, University of Michigan, School of Medicine, 7730 Medical Science II, Ann Arbor, MI 48109-0622.
Abstract 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 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–polyacrylamide gel electrophoresis. The distribution
of MHC isoform expression in isolated ES cell cardiac myocytes was as
follows: 27% expressed the ß-MHC isoform, 33% expressed both the
- and ß-MHC isoforms, and 40% expressed the -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 ß-MHC (average days
of contraction before isolation, 2.5±0.7). The -MHC isoform was
detected after more 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 ß- to
-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.
Key Words: contraction • development • cardiac muscle
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