低密度和条件培养对红豆杉细胞生长的影响

Effects of Low Density Cell Culture and Conditioned Culture of Taxus chinensis Cells in Suspension-cultures

红豆杉种胚来源的细胞 ,在改良 B5液体培养基中继代培养的临界接种密度为鲜重 40g/L .低密度培养下 ,1 0～ 1 6d的条件培养液 ( CM)与新鲜培养液按 57∶ 43的比例混合时 ,能显著缩短细胞生长的延迟期 ,提高生长率 .1 0 0 L生物反应器中 ,按 45.5%体积分数添加条件培养液 ,在鲜重 2 7g/L低接种密度下培养 5周 ,生物量增长 9倍 ,达干重 1 4.3g/L.对内源植物激素、精胺、维生素和氨基酸等的比较分析表明 ,吲哚乙酸和精胺在低密度培养下可在一定程度上替代条件培养液 .两者协同作用 ,使细胞生物量增长率较常规接种密度时提高约 2倍 .

Low density cell culture and conditioned culture of Taxus chinensis cells suspended in flasks and 100 litre stirred tank bioreactors were studied. When the cells were cultured in a growth medium (modified B 5 medium), the subculture critical inoculum density of T. chinensis was 40 g/L (fresh weight). Conditioned medium (CM) extracted from yew suspension cultures between 10 to 16 days after inoculation to fresh medium at a terminal concentration of volume fraction 57.1% resulted faster cell growth than in low density cell culture [4 g/L (fresh weight)]. Similar effect was found by applying indoleacetic acid (IAA) and spermine but not by other compounds (including amino acids, vitamins and zeatin). When IAA and spermine were used in combination, the routine inoculum density of T. chinensis suspension cells could be reduced from 100 g/L (fresh weight) to 40 g/L (fresh weight). Yew cells cultured in 100 litre STB for thirty six days gave a 9 fold dry weight increase gaining a biomass concentration of 14.3 g/L (dry weight), when CM was mixed with fresh medium in the ratio of 45.5 to 54.5 at an inoculum density of 27 g/L (fresh weight). [WT5HZ]