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取材方法对培养的新生大鼠心室细胞电生理及形态学特性的影响。

Influence of explantation procedure on the electrical and morphological properties of cultured neonatal rat ventricle cells.

作者信息

Fermini B, Schanne O F, Hugon J S

出版信息

Adv Myocardiol. 1985;6:25-36.

PMID:2986259
Abstract

The ultrastructure and electrophysiological properties of ventricle cells from newborn rats were studied before and after explantation. The cultured cells were dissociated either with trypsin or with collagenase, the latter enzyme being used with and without stirring with a magnetic bar. The explanted cells were studied 10 hr and 48 hr or more after explantation. At 10 hr after explantation, the cells exhibited fast-rising action potentials, but their myofibrils were disorganized, except for stirred collagenase-dispersed cells, which were also depolarized and inexcitable. At 2 days and later after explantation, all preparations had well-defined sarcomeres and myofibrils oriented in parallel similar to the ventricle before explantation, but the cells showed slow-response action potentials together with spontaneous activity. These findings suggest that the disorganization of myofibrils does not reflect damage to the surface membrane. Moreover, collagenase seems more damaging to the cells than trypsin under similar conditions (comparable periods of mechanical stirring), especially 10 hr after explantation.

摘要

对新生大鼠心室细胞在植入前和植入后的超微结构及电生理特性进行了研究。培养的细胞用胰蛋白酶或胶原酶解离,后者在有或没有磁棒搅拌的情况下使用。植入后10小时和48小时或更长时间对植入的细胞进行研究。植入后10小时,细胞表现出快速上升的动作电位,但其肌原纤维排列紊乱,除了经搅拌的胶原酶分散的细胞,这些细胞也发生了去极化且无兴奋性。植入后2天及更晚,所有标本都有明确的肌节,肌原纤维平行排列,类似于植入前的心室,但细胞表现出慢反应动作电位以及自发活动。这些发现表明,肌原纤维的紊乱并不反映表面膜的损伤。此外,在类似条件下(机械搅拌时间相当),胶原酶对细胞的损伤似乎比胰蛋白酶更大,尤其是在植入后10小时。

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