怀牛膝悬浮细胞培养的动力学研究

Kinetics in Suspension Cell Culture of Achyranthes bidentata Bl.

以怀牛膝悬浮细胞系为试材,对牛膝细胞悬浮培养动力学进行了研究,采用HPLC、原子吸收分光光度法和电导率测定等方法,分析了牛膝悬浮细胞生长、营养消耗和三萜皂苷积累的动力学关系。结果表明:牛膝细胞悬浮培养周期约为24d,经24d的悬浮培养,最大生物量和齐墩果酸含量分别达到了13.47mg/L和1.418mg/g;牛膝悬浮细胞生长与三萜皂苷积累的动力学关系属于生长偶联型。培养过程中培养液的pH值呈先下降后回升的趋势,而电导率逐渐减小,24d时降到最低点,而后略有上升。经15d培养,培养基中的蔗糖即消耗殆尽;培养21d后,培养基中大量的氮源、磷源、K+和Mg2+被消耗,而Ca2+比较充足,基本可以满足细胞生长的需要。由此可推断,培养液中营养成分的不足限制了细胞的快速增长和三萜皂苷含量的提高。

Taking the cell culture of Achyranthes bidentata Bl.as material,using HPLC,atomic absorption spectrometry method and conductivity measurement,the kinetic relationships of suspension cell growth,triterpenoid saponins production and nutrient consumption were analyzed.The results showed the suspension cell culture cycle lasted about 24 days by which the maximum biomass in dry weight and the oleanolic acid content reached to 13.47mg/L and 1.418mg/g.The kinetics analysis for suspension cell culture of A.bidentataBl.showed that triterpenoid saponins production was increasing with the cell growth during a cell subculture cycle.The electrical conductivity of the culture medium gradually descended during culture procedure,reached to the lowest point on the 24 th day,and then slightly increased.The pH value declined and then increased.After 15 days of the cell culture,sucrose were consumed in culture medium.The large amounts of nitrogen,phosphate,K＋and Mg2＋were consumed on the 21 st days.Compared with K＋,Ca2＋could meet the needs of the cell growth.The results suggested that the nutrient deficiencies in medium would limite the cell rapid multiplication and triterpenoid saponins content accumulation.