Myosin heavy chain expression in contracting myocytes isolated during embryonic stem cell cardiogenesis

Joseph M. Metzger, Wan In Lin, Ross A. Johnston, Margaret V. Westfall, Linda C. Samuelson

Research output: Contribution to journalArticle

63 Scopus citations

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 β-MHC isoform, 33% expressed both the α- and β-MHC isoforms, and 40% expressed the α-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 β-MHC (average days of contraction before isolation, 2.5±0.7). The α-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 β- to α-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.

Original languageEnglish (US)
Pages (from-to)710-719
Number of pages10
JournalCirculation Research
Volume76
Issue number5
DOIs
StatePublished - May 1995

Keywords

  • cardiac muscle
  • contraction
  • development

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