| 摘要: |
| 金钱鱼(Scatophagus argus)是一种重要的广盐性海水养殖鱼类, 可直接在海水、淡水、咸淡水等不同环境中正常生长。为研究金钱鱼独特的渗透调节机制, 本文研究了金钱鱼肾细胞的原代和传代条件及其生长特性, 结果表明: 原代肾细胞在含有20%胎牛血清(FBS)的L-15 培养基里贴壁和生长较好, 添加10 ng/mL 的碱性成纤维细胞因子(basic fibroblast growth factor) bFGF 能明显促进细胞增殖, 传代细胞采用含有10%胎牛血清的L-15 培养基, 细胞生长迅速, 3-4d 即可传代。金钱鱼肾细胞为成纤维样细胞, 命名为SK 细胞, 目前已传至22 代。扩增第11 代SK 细胞的细胞色素氧化酶I(COI)基因, 比对结果证明此细胞系来源于金钱鱼。采用CCK-8 法检测第12 代细胞在低渗(95, 137,200 mmol/kg), 等渗(330 mmol/kg) 和高渗(430, 550 mmol/kg) 中的增殖情况, 结果发现肾细胞在137-430 mm/kg 的渗透压范围内均可增殖, 说明金钱鱼肾细胞对渗透压的耐受性较强。本试验首次建立了金钱鱼肾细胞系, 并初步证明了肾细胞对盐度有较强的耐受性, 为今后渗透压调节的深入研究奠定了基础。 |
| 关键词: 金钱鱼 肾细胞系 原代培养 传代培养 渗透压调节 |
| DOI：10.11693/hyhz20130131001 |
| 分类号: |
| 基金项目:国家科技部 “十二五”“科技支撑计划”, 2011BAD13B00 号; 国家自然科学基金项目, 41176109 号 |
附件 |
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| ESTABLISHMENT AND CHARACTERIZATION OFA NEW EURYHALINE FISH KIDNEY CELL LINE OF SPOTTED SCAT SCATOPHAGUS ARGUS |
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zhang jun bin and zhang ying ying
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Shanghai Ocean University,Shanghai Ocean University
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| Abstract: |
| Spotted scat Scatophagus argus is a euryhaline fish and they could live in seawater, freshwater, or brackish water. We established and characterized a new kidney cell line (SK) of the fish, and successfully subcultured SK to the 22th generations at present. The results showed that the primary cells grew well in L-15 medium with 20% fetal bovine serum (FBS), and adding 10 ng/mL basic fibroblast growth factor (bFGF) could increase significantly the growth rate. Subcultured cells were maintained in L-15 medium with 10% FBS and could be passaged in 3-4 days. SK cells were observed fibroblastic. PCR amplification of l cytochrome oxidase subunit I (COI) for the cells of 11th generation confirmed the origin of this cell line. The CCK-8 method was used to detect the proliferation rate for the twelfth SK cells in hypotonic (95, 137, 200 mmol/kg), isotonic (330 mmol/kg), and hypertonic (430, 550 mmol/kg) conditions. Therefore, the SK cells were able to proliferate in the wide range from 137 to 430 mm/kg osmotic pressure. |
| Key words: Scatophagus argus kidney cell line primary culture subculture osmoregulation |
