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为了建立条斑星鲽鳍细胞系,为其细胞工程及病毒学研究奠定基础,对经Ⅱ型胶原酶和透明质酸酶联合消化法所得鳍组织碎块分别用DMEM/F12、L-15和M199培养液(pH7.2)在18～26℃进行了鳍组织的体外培养,并在所筛选出的最适培养液和培养温度下通过添加羧甲基壳寡糖、碱性成纤维样生长因子(bFGF)和I型胰岛素样生长因子(IGF-I)启动了鳍细胞的原代培养。体外培养结果显示,条斑星鲽鳍细胞的最适培养液为DMEM/F12培养液,最适培养温度为22℃,原代培养鳍细胞的生长分裂状态旺盛,细胞形态主要为成纤维样,20d后便可形成汇合细胞单层,经过连续的继代培养已建立了条斑星鲽的连续性鳍细胞系,目前已传至第135代;生长特性检测结果显示,第60代鳍细胞系细胞的群体倍增时间为56.9h,其生长分裂状态依然十分旺盛;染色体分析结果显示,第60代鳍细胞系细胞虽然出现了染色体的非整倍性,但其特征性染色体数目仍为46条,并具有2sm+44t的正常二倍体核型,证明所建立的细胞系确为条斑星鲽连续性鳍细胞系。该细胞系为条斑星鲽的细胞工程育种和病毒-细胞相互作用提供了一个理想的体外研究体系,具有重要的理论意义和应用价值。
In order to establish the streak-star-shaped fin fins cell line, which laid the foundation for its cellular engineering and virology research, the fragments of fin tissue obtained by digestion of type II collagenase and hyaluronidase were respectively treated with DMEM / F12, L-15 and M199 culture broth (pH7.2) was cultured in vitro at 18 ~ 26 ℃ for fin tissue, and basic fibroblast-like growth was carried out by adding carboxymethyl chitooligosaccharides at the selected optimal medium and culture temperature Primary culture of fin cells is initiated by bFGF and IGF-I. The results of in vitro culture showed that the optimal culture medium for the cells of Sturnus striatus was DMEM / F12, the optimal culture temperature was 22 ℃, the primary cultured fin cells had strong growth and division status, the main cell morphology was fibroblast-like, After 20 days, a single layer of confluent cells could be formed. After continuous subculture, a continuous fin cell line was established, which has been transmitted to the 135th generation. The growth characteristics test results showed that the 60th generation fin cell line The population doubling time was 56.9h and its growth and division status was still very strong. The results of chromosome analysis showed that the number of chromosomes in the 60th-generation fins cell lines were still 46 aneuploidy, And has a normal diploid karyotype of 2sm + 44t, demonstrating that the established cell line is indeed a streak-feeding fin fins cell line. This cell line provides an ideal in vitro research system for cell engineering breeding and virus-cell interaction of St. Osprey and has important theoretical significance and application value.