改良的L-15培养基对鳞翅目昆虫细胞生长和芹菜夜蛾核型多角体病毒增殖的影响

Growth of Some Lepidopteran Cell Lines in Modified L-15 Media and Replication of Anagrapha falcifera Multiple Nucleopolyhedrovirus in the Cells

4种鳞翅目昆虫细胞系SL-ZSU-1、TnH5、Sf9和Ha．AM1都不能在L-15＋5％FBS或L-15＋350mg／L脯氨酸＋5％FBS（L-15-P，pH6．5）的培养基中生长，但是都可以在L-15＋3g／L水解乳蛋白＋3g／L酵母提取物＋1g／L葡萄糖＋5％FBS（L-15-LYG）、L．15＋350mg／L脯氨酸＋1g／L葡萄糖＋5％FBS（L-15-PG）、L-15＋1g／L葡萄糖＋5％FBSa（L-15-G，pH6．5）的培养基中生长。细胞从TNM-FH＋5％FBS（pH6．5）培养基中转入上述培养基后，适应期的长短相差较大。转入L-15-LYG中的细胞没有明显的适应期；转入L-15-PG中的细胞需1个月左右才能正常传代；转入L-15-G中的细胞需45～60d才能正常传代。L-15-LYG替代TNM-FH对4种昆虫细胞的形态、大小、最大生长密度和群体培增时间没有显著影响，对芹菜夜蛾核型多角体病毒AfaMNPV胞外病毒粒子的滴度没有明显的变化，但多角体的产量显著下降。

The tested four lepidopteran cell lines （SL-ZSU-1, Tn-HS, Sf9 and Ha-AM1 ） can not grow in L-15 ＋ 5% FBS （L-15）, or L-15 ＋ 350mg/L proline ＋ 5% FBS （L-15-P） at pH 6.5, but they can grow in L-15 medium supplemented with lg/L glucose and 5%FBS （L-15-G）, L-15 ＋ 350mg/L proline ＋ lg/ Lglucose ＋ 5%FBS （L-15-PG）, or L-15 ＋ 3g/L lactalbumin hydrolysate ＋ 3g/L yeast extract ＋ lg/L glu- cose ＋ 5 % FBS （ L- 15- LYG）. The time needed for adaptation to these modified L- 15 media was different. The four cell lines could be rapidly adapted to L-15-LYG medium, but the time needed for adaptation to L-15-PG and L-15-G was one month and 45 - 60d, respectively. The growth of the four cell lines in L- 15-LYG medium was equal to that in TNM-FH medium. The titers of Anagraphafalcifera multiple nucleopolyhedrovirus （AfaMNPV） for Tn-H5 and Sf9 cell lines cultured in L-15-LYG medium were as good as that in TNM-FH medium, but the production of occlusion bodies （OBs） was decreased.