Kinetic properties of extracted lactate dehydrogenase and creatine kinase from mouse embryonic stem cell- and neonatal-derived cardiomyocytes.
Embryonic stem cells (ESCs), representing a population of undifferentiated pluripotent cells with both self-renewal and multilineage differentiation characteristics, are capable of spontaneous differentiation into cardiomyocytes. The present study sought to define the kinetic characterization of lactate dehydrogenase (LDH) and creatine kinase (CK) of ESC- and neonatal-derived cardiomyocytes. Spontaneously differentiated cardiomyocytes from embryoid bodies (EBs) derived from mouse ESC line (Royan B1) and neonatal cardiomyocytes were dispersed in a buffer solution. Enzymes were extracted by sonication and centrifugation for kinetic evaluation of LDH and CK with spectrophotometric methods. While a comparison between the kinetic properties of the LDH and CK of both groups revealed not only different Michaelis constants and optimum temperatures for LDH but also different Michaelis constants and optimum pH for CK, the pH profile of LDH and optimum temperature of CK were similar. In defining some kinetic properties of cardiac metabolic enzymes of ESC-derived cardiomyocytes, our results are expected to further facilitate the use of ESCs as an experimental model.
Zonouzi
Roseata
R
Department of Stem Cells, Royan Institute, Tehran, Iran.
Ashtiani
Saeid Kazemi
SK
Hosseinkhani
Saman
S
Baharvand
Hossein
H
D000818:Animals; D000831:Animals, Newborn; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D002469:Cell Separation; D003402:Creatine Kinase / Q000378:metabolism*; D004622:Embryo, Mammalian / Q000166:cytology; D007700:Kinetics; D045022:Lactate Dehydrogenases / Q000378:metabolism*; D051379:Mice; D032383:Myocytes, Cardiac / Q000166:cytology* / Q000201:enzymology*; D013234:Stem Cells / Q000166:cytology*
D045022:Lactate Dehydrogenases; D003402:Creatine Kinase