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[新生大鼠心室细胞培养:DNA合成、超微结构及心房利钠肽的定位]

[Heart ventricular cells of neonatal rats in a culture: DNA synthesis, ultrastructure and localization of atrial natriuretic peptide].

作者信息

Erokhina I L, Semenova E G, Emel'ianova O I, Krylova M I

机构信息

Institute of Cytology RAS, St. Petersburg.

出版信息

Tsitologiia. 2003;45(7):621-7.

Abstract

We determined the optimal conditions suitable for expanding cardiac cells in vitro for their future use in experimental transplantation into injured myocardium of adult animals. Ventricular cardiac cells were isolated enzymatically from 2-3 day-old rats and cultured at different cell densities within 5-7 days to 4 weeks. Mixed cultures of muscle and non-muscle cells were examined by light autoradiography, electron microscopy, and immunogold method. The best results were obtained at a density of 3 x 10(5) cells/ml in the medium, consisting of 90% DMEM and 10% fetal calf serum, during 5-7 days of cultivation. In such cultures myocytes made 62.5 +/- 7.9%. After a 24 h incubation with 3H-thymidine, 22.0 +/- 2.2% of myocytes were labeled. Muscle cells contact with each other and with non-muscle cells, contain myofibrils, contract and display atrial natriuretic peptide (ANP)-like immunoreactivity.

摘要

我们确定了适合体外扩增心脏细胞的最佳条件,以便将来用于实验性移植到成年动物受损心肌中。从2 - 3日龄大鼠中酶法分离心室心肌细胞,并在不同细胞密度下培养5 - 7天至4周。通过光镜放射自显影、电子显微镜和免疫金法检查肌肉和非肌肉细胞的混合培养物。在由90% DMEM和10%胎牛血清组成的培养基中,细胞密度为3×10(5) 个细胞/ml,培养5 - 7天时获得了最佳结果。在这种培养物中,心肌细胞占62.5±7.9%。用3H - 胸苷孵育24小时后,22.0±2.2%的心肌细胞被标记。肌肉细胞相互接触并与非肌肉细胞接触,含有肌原纤维,能收缩并显示出心房利钠肽(ANP)样免疫反应性。

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