| 摘要: |
| 采用改良的DMEM(HG)培养基, 建立了近江牡蛎(Crassostrea hongkongensis)鳃细胞体外培养技术, 包括鳃组织块培养法和胰酶消化培养法。结果表明, 组织块培养法接种6 h 后, 细胞开始从组织块中迁出, 细胞形态较小, 呈圆形、椭圆形或多边形, 直径3～6 μm。培养至3 d 时, 细胞在组织块周围形成生长晕。培养至6 d 时可进行细胞传代, 本次实验细胞已传至第6 代。胰酶消化法接种约2 h 后, 细胞逐渐贴壁, 从形态上主要分为两类, 一类为小型细胞, 形态为圆形、椭圆形或多边形, 直径3～6 μm,数量多, 增殖速度较快; 另一类为大型细胞, 形态为圆形、椭圆形或多边形, 直径10～20 μm, 部分细胞内部含有颗粒, 数量较少, 增殖速度较慢。培养至2 d 时可进行细胞传代, 传代培养物中的优势细胞皆为小型细胞。本次实验细胞已传至第6 代。 |
| 关键词: 近江牡蛎(Crassostrea hongkongensis) 鳃细胞 组织块培养 胰酶消化 |
| DOI： |
| 分类号: |
| 基金项目:国家自然科学基金项目(40576056, 40976066)资助 |
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| Primary culture of oyster (Crassostrea hongkongensis) gill cells |
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WANG Bin,ZHANG Qi-zhong
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| Abstract: |
| The gill cells derived from oyster (Crassostrea hongkongensis) were cultured in vitro with the modified medium DMEM (HG) by means of both gills tissue culture and trypsinization methods. In the gills tissue culture method, the small cells, which were rounded, elliptic or polygonal with diameters of 3～6 μm, began to migrate out from the gills tissue at 6 h post-inoculation. The cell layers appeared around the tissues at 3 d post-inoculation, and could be subcultured at 6 days. In present study, the gill cells had been subcultured to the sixth generation. In the trypsinization method, most gill cells were adherent at 2 h post-inoculation of trypsinization. The cells could be morphologically divided into two groups including small cells with diameters of 3～6 μm and large cells with diameters of 10～20 μm. The two groups of cells were all rounded, elliptic or polygonal; some of the large cells were granular cells. The small cells were more than the large cells in number. Being cultured for 2 d, the gill cells could be subcultured. In the present study, the gill cells had been subcultured to the sixth generation. |
| Key words: Crassostrea hongkongensis gill cells tissue culture trypsinization |
