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从斑点叉尾鮰中分离、培养和鉴定原代成纤维细胞系。

Isolation, culture and characterization of a primary fibroblast cell line from channel catfish.

机构信息

School of Forestry, Wildlife, and Fisheries, Louisiana Agricultural Experiment Station, Louisiana State University Agricultural Center, Baton Rouge, LA, 70803, USA.

出版信息

Cytotechnology. 1998 Mar;26(2):83-90. doi: 10.1023/A:1007911619537.

Abstract

A primary cell line (designated as CCf) derived from caudal fin tissue of channel catfish, Ictalurus punctatus, was developed using explant techniques. The cell line grew fastest in media supplied with FBS and channel catfish serum. The duplication time of the cell line under optimal conditions was ∼56 h at a plating density of 1.1 × 10(5) cells/ml. The cell line has been propagated continuously for 25 passages (1:4 dilution per passage), cryopreserved, and recovered successfully at different passages. The cultured cells had fibroblastic morphology, and synthesized fibronectin and Type I and III collagens in the cytoplasm. The cell line maintained the normal diploid chromosome number (58) of channel catfish throughout the experiment. Nucleolus organizer regions were located on the short arms of a pair of medium-sized submetacentrics, which is typical for channel catfish. This study provides a method for acquiring a cell line from juvenile catfish without sacrifice, and is especially useful for early screening of valuable fishes.

摘要

本研究采用组织块培养法,从美洲鲶(Ictalurus punctatus)尾鳍组织中成功建立了一个原代细胞系(命名为 CCf 细胞系)。该细胞系在含胎牛血清和美洲鲶血清的培养基中生长最快。在最优条件下,细胞系的倍增时间约为 56 小时,接种密度为 1.1×10(5)细胞/ml。该细胞系已连续传代 25 次(每传代稀释 1:4),并成功进行了冷冻保存和复苏。培养的细胞呈成纤维细胞形态,在细胞质中合成纤连蛋白和 I 型和 III 型胶原蛋白。在整个实验过程中,细胞系保持了美洲鲶正常的二倍体染色体数目(58 条)。核仁组织区位于一对中等大小的亚中部着丝粒的短臂上,这是美洲鲶的典型特征。本研究提供了一种从幼鱼中获取细胞系而无需牺牲的方法,特别适用于早期筛选有价值的鱼类。

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