匙吻鲟鳍条组织细胞系的建立及其生物学特性

Establishment and Characterization of a Cell Line Derived From Fin of Paddlefish,Polyodon spathula Walbaum

采用组织块移植培养技术,对来源于匙吻鲟鳍条组织的细胞进行原代培养,建立了匙吻鲟鳍条组织细胞系,已稳定传代培养80多代,定名为PF-Fin。匙吻鲟鳍条组织细胞的形态为成纤维样细胞,其最佳培养基为M199,最适血清浓度为10%,最适培养温度为25℃。液氮冷冻保存8个月后的细胞经台盼兰染色检验,约87.68%±3.61%仍保持活性,细胞复苏后培养生长旺盛。匙吻鲟鳍条组织细胞的集落形成效率为13.75%±2.28%;细胞染色体分析结果表明,第10代传代细胞的染色体数目为正常二倍体2n=120,第59代传代培养细胞的染色体众数为90。病毒敏感性试验结果表明,PF-Fin细胞对草鱼呼肠孤病毒(GCRV)、斑点叉尾鮰呼肠孤病毒(CCRV)和鲤鱼春季病毒血症病毒(SVCV)敏感,可产生典型细胞病变效应(CPE)。

A cell line, designated PF-Fin, had been established from the fin of paddlefish, Polyodon spathula Walbaum and subcultured over 80 passages. Characterization of the cell line included determination of cell morphology, optimal growth kinetics, plating efficiency, karyotyping, etc. The primary culture of this cell was generated by the tissue explant technique using the M199 medium supplemented with 30% fetal bovine serum, epidermal/fibroblast growth factors and penicillin/streptomycin/-amphotericin B solution. Subculture of the cell was conducted in M199 medium with 10% fetal bovine serum. The cell grew between 20-30 ℃ and the optimal growth temperature was 25 ℃. The cell monolayer consisted of fibroblast-like cells and the plating efficiency of the cells was about 13.75%±2.28%. Following cryopreservation in liquid nitrogen, thawed cells exhibited a viability of 87.68%＋3.61% after 8 months storage period. Chromosome typing of this cell line revealed the diploid chromosome number was 2n=120 at its 10th passage, and at its 59th passage the chromosome modal number was 2n=90. The results of virus challenge tests indicated that PF-Fin cell line is susceptible to the infection of Grass carp Reovirus （GCRV）, Channel catfish Reovirus （CCRV） and Spring Viriemia carp Virus （SVCV）, and a characteristic viral-induced cytopathic effect （CPE） was observed.