Schackow T E, Sheets M F, Decker R S, Ten Eick R E
Department of Molecular Pharmacology and Biological Chemistry, Northwestern University, Chicago, Illinois 60611, USA.
Am J Physiol. 1995 Apr;268(4 Pt 1):C993-1001. doi: 10.1152/ajpcell.1995.268.4.C993.
To determine the response of cardiac Na current (INa) in adult cardiac ventricular myocytes to culture, single isolated ventricular myocytes from collagenase-perfused adult cat hearts were placed in primary culture for up to 2 wk on a two-dimensional (2D) surface (laminin-coated coverslips), which allowed the morphology of the myocytes to change markedly, or in a three-dimensional matrix (3D) of alginate, in which cell shape changed only minimally. Action potentials and INa were recorded from groups of 1) freshly isolated myocytes serving as the control (day 0),2) cells maintained in 2D culture for 9-14 days (2D, day 9-14), and 3) cells cultured in alginate for 9-14 days (3D, day 9-14) with use of a conventional whole cell patch technique. Maximal upstroke velocity (Vmax) of the action potential was reduced by approximately 50% in 2D- and 3D-cultured cells relative to controls. INa in 2D- and 3D-cultured cells was strikingly different from that in control myocytes. Half-maximal voltage (V 1/2) for the chord conductance-voltage relationship was shifted approximately 15 mV negatively to that for controls in 2D- and 3D-cultured cells. INa steady-state availability curve also shifted negatively relative to controls in 2D- and 3D-cultured myocytes, but the magnitude of this shift (approximately 16-20 mV) was greater than that for the chord conductance-voltage curve.(ABSTRACT TRUNCATED AT 250 WORDS)
为了确定成年心脏心室肌细胞中的心钠电流(INa)对培养的反应,将来自胶原酶灌注成年猫心脏的单个分离心室肌细胞在二维(2D)表面(层粘连蛋白包被的盖玻片)上进行原代培养长达2周,这会使肌细胞形态发生显著变化,或者在藻酸盐三维基质(3D)中培养,其中细胞形状变化极小。使用传统的全细胞膜片技术从以下几组细胞中记录动作电位和INa:1)作为对照的新鲜分离肌细胞(第0天),2)在2D培养中维持9 - 14天的细胞(2D,第9 - 14天),以及3)在藻酸盐中培养9 - 14天的细胞(3D,第9 - 14天)。相对于对照,2D和3D培养细胞的动作电位最大上升速度(Vmax)降低了约50%。2D和3D培养细胞中的INa与对照肌细胞中的显著不同。2D和3D培养细胞中弦电导 - 电压关系的半数最大电压(V1/2)相对于对照向负向移动了约15 mV。在2D和3D培养的心肌细胞中,INa稳态可用性曲线相对于对照也向负向移动,但这种移动的幅度(约16 - 20 mV)大于弦电导 - 电压曲线的移动幅度。(摘要截断于250字)