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离子通道在克隆肾细胞体积调节中的作用。

Ion channels in volume regulation of clonal kidney cells.

机构信息

Department of Biophysics and Radiobiology, Federal University of Pernambuco, Recife, PE, Brazil.

出版信息

Cell Prolif. 2010 Dec;43(6):529-41. doi: 10.1111/j.1365-2184.2010.00702.x.

Abstract

OBJECTIVES

Clonal kidney cells (Vero cells) are extensively utilized in the manufacture of biological preparations for disease diagnostics and therapeutics and also in preparation of vaccines. In all cells, regulation of volume is an essential function coupled to a variety of physiological processes and is a topic of interest. The objective here was to investigate involvement of ion channels in the process of volume regulation of Vero cells.

METHODS

Involvement of ion channels in cell volume regulation was studied using video-microscopy and flow cytometry. Pharmacologically unaltered cells of different sizes, which are presumably at different phases of the cell cycle, were used.

RESULTS

Ion transport inhibitors altered all phases of regulatory volume decrease (RVD) of Vero cells, rate of initial cell swelling, V(max) and volume recovery. Effects were dependent on type of inhibitor and on cell size (cell cycle phase). Participation of aquaporins in RVD was suggested. Inhibitors decelerated growth, arresting Vero cells at the G(0) /G(1) phase boundary. Electrophysiological study confirmed presence of volume-activated Cl(-) channels and K(+) channels in plasmatic membranes of the cells.

CONCLUSION

Vero cells of all sizes maintained the ability to recover from osmotic swelling. Activity of ion channels was one of the key factors that controlled volume regulation and proliferation of the cells.

摘要

目的

克隆肾细胞(Vero 细胞)广泛应用于疾病诊断和治疗的生物制剂生产,也应用于疫苗制备。在所有细胞中,体积调节是与多种生理过程相关的基本功能,是一个研究热点。本研究旨在探讨离子通道在 Vero 细胞体积调节过程中的作用。

方法

使用视频显微镜和流式细胞术研究离子通道在细胞体积调节中的作用。使用不同大小的、可能处于不同细胞周期阶段的未经药物处理的细胞。

结果

离子转运抑制剂改变了 Vero 细胞的所有调节性体积减少(RVD)阶段、初始细胞肿胀的速度、V(max)和体积恢复。这些作用取决于抑制剂的类型和细胞大小(细胞周期阶段)。提示水通道蛋白参与了 RVD。抑制剂减缓了细胞生长,将 Vero 细胞阻滞在 G0/G1 期边界。电生理学研究证实细胞质膜中存在体积激活的 Cl(-)通道和 K(+)通道。

结论

所有大小的 Vero 细胞均保持从渗透肿胀中恢复的能力。离子通道的活性是控制细胞体积调节和增殖的关键因素之一。

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