曼地亚红豆杉细胞系的建立与评价

Establishment and Assessment of Taxus media Cell Line

红豆杉(Taxus)细胞内紫杉醇的含量低且不稳定,限制了利用细胞培养大规模生产紫杉醇的产业化进程。以紫杉醇含量较高的曼地亚红豆杉(Taxus m edia)为试材,诱导得到了曼地亚红豆杉愈伤组织,在添加抗坏血酸(VC)、聚乙烯吡咯烷酮(PVP)、活性炭(activated carbon)的改良B5培养基上,15次反复继代培养后获得了高产紫杉醇的细胞系。对新建立的曼地亚红豆杉细胞系(MC)与同期诱导的东北红豆杉细胞系(NC)及实验室继代多年的中国红豆杉细胞系(SC)在细胞生长周期、紫杉醇含量和细胞死亡率等方面进行了分析比较。结果表明,SC的生长量达5.9倍,高于MC和NC的3.6和4.2倍。在初始接种量相近的情况下,MC的悬浮培养细胞在一个周期内紫杉醇产量可达9.5 mg/L,经过茉莉酸甲酯(M J)诱导后可达到41 mg/L。

Taxol in Taxus cell is little and unstable, which prevents the industrialization process of producing taxol by cell culture. Using Taxus med/a as material, the T. media callus was accrued. After subculturing on solid modified B5 medium for 15 time, and supplied with VC, PVP and activated carbon, cell line with the ability of high taxol production was attained. Cell growth cycle, taxol content and cell death rate of T. media（ MC）, T. cuspidate（NC） which was successfully established at the same time and T. chinensis （SC） which was subcuhured for several years in the lab was studied. The results show that the biomass of SC reaches the peak of 5.9 times, which is higher than that of NC and MC. With the same initial inoculation weight, taxol production of MC reached at 9.5 mg/L, and that could reach at 41mg/L after induced by MJ.