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从小鼠胚胎干细胞和新生来源的心肌细胞中提取的乳酸脱氢酶和肌酸激酶的动力学特性。

Kinetic properties of extracted lactate dehydrogenase and creatine kinase from mouse embryonic stem cell- and neonatal-derived cardiomyocytes.

作者信息

Zonouzi Roseata, Ashtiani Saeid Kazemi, Hosseinkhani Saman, Baharvand Hossein

机构信息

Department of Stem Cells, Royan Institute, Tehran, Iran.

出版信息

J Biochem Mol Biol. 2006 Jul 31;39(4):426-31. doi: 10.5483/bmbrep.2006.39.4.426.

Abstract

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.

摘要

胚胎干细胞(ESC)是一群具有自我更新和多谱系分化特征的未分化多能细胞,能够自发分化为心肌细胞。本研究旨在确定胚胎干细胞来源的心肌细胞和新生心肌细胞中乳酸脱氢酶(LDH)和肌酸激酶(CK)的动力学特征。从小鼠胚胎干细胞系(Royan B1)来源的胚状体(EB)自发分化得到的心肌细胞以及新生心肌细胞被分散在缓冲溶液中。通过超声处理和离心提取酶,采用分光光度法对LDH和CK进行动力学评估。虽然两组LDH和CK的动力学特性比较显示,不仅LDH的米氏常数和最适温度不同,而且CK的米氏常数和最适pH也不同,但LDH的pH曲线和CK的最适温度相似。在确定胚胎干细胞来源的心肌细胞心脏代谢酶的一些动力学特性时,我们的结果有望进一步促进将胚胎干细胞用作实验模型。

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