Mitcheson J S, Hancox J C, Levi A J
Department of Physiology, University of Bristol, School of Medical Sciences, UK.
Cardiovasc Res. 1998 Aug;39(2):280-300. doi: 10.1016/s0008-6363(98)00128-x.
Isolated adult cardiac myocytes maintained in primary culture have been used as a model of the adult myocardium for 20 years. With the recent advances and current interest in using molecular biological techniques to investigate cardiac physiology, culturing myocytes is becoming an increasingly important technique. Acutely isolated myocytes do not remain viable for the time needed for the changes in gene expression to occur, and therefore it is necessary to maintain myocytes in culture. The aims of this review are: (1) To describe a method for isolating and culturing myocytes in serum-free medium. This section is targeted at new researchers in the field, with particular emphasis on aspects of the isolation procedure which are important for optimising myocyte culture. (2) To review current knowledge of how contractile, electrophysiological and morphological properties of adult myocytes are preserved in culture. Over the past 5 to 10 years significant advances have been made in developing novel techniques which help maintain the in-vivo properties of myocytes in culture. Efficient methods for transporting exogenous genes and anti-sense oligonucleotides into adult myocytes are now available. We anticipate that in future these advances will make cultured myocytes more attractive for use in biophysical and molecular investigations of cardiac physiology.
原代培养的成年离体心肌细胞作为成年心肌模型已被使用了20年。随着近年来在运用分子生物学技术研究心脏生理学方面取得的进展以及当前对该领域的关注，培养心肌细胞正成为一项日益重要的技术。急性分离的心肌细胞无法在基因表达发生变化所需的时间内保持存活，因此有必要将心肌细胞进行培养。本综述的目的如下：（1）描述一种在无血清培养基中分离和培养心肌细胞的方法。此部分针对该领域的新研究人员，特别强调分离过程中对优化心肌细胞培养至关重要的方面。（2）综述目前关于成年心肌细胞的收缩、电生理和形态学特性在培养中如何得以保留的知识。在过去5到10年里，在开发有助于在培养中维持心肌细胞体内特性的新技术方面取得了重大进展。现在已有将外源基因和反义寡核苷酸有效导入成年心肌细胞的方法。我们预计，未来这些进展将使培养的心肌细胞在心脏生理学的生物物理和分子研究中更具吸引力。
