红豆杉细胞植板方法的研究

在红豆杉细胞植板中，适宜的培养基是优化的ＭＳ培养基，其中ＮＡＡ浓度为０．５ｍｇ／Ｌ，６ＢＡ浓度为０．５ｍｇ／Ｌ；培养细胞生长周期约４０ｄ；培养基中最优碳源浓度是：蔗糖３０ｇ／Ｌ、葡萄糖１０ｇ／Ｌ．无机盐最佳浓度是：ＮＨ＋４１０ｍｍｏｌ／Ｌ，ＮＯ－３４０ｍｍｏｌ／Ｌ，ＰＯ３－４１．２５ｍｍｏｌ／Ｌ．研究表明，培养基的ｐＨ值与植板率密切相关．氨基酸及维生素等有机成分能明显促进细胞生长．来自细胞悬浮培养物的条件培养基能显著地促进红豆杉培养细胞的单细胞在低密度植板时的克隆形成

On the Method of Plating Cell Culture from Taxus Chinensis

In callus plate of taxus,the most suitable medium was the modified MS medium added 0.5mg/L NAA and 0.5mg/L 6BA. The growth curve of cell exhibited that the optimum period for callus subculture was 40 days; The best combination of sucrose and glucose was 30g/L and 10g/L respectively. The fittest concentration of NH + 4,NO - 3 and PO 3- 4 were 10mmol/L,40mmol/L and 1.25mmol/L respectively. The optimum pH was 5.8 for cell plating culture. The plating efficiency could also be effectively increased when the plating medium was suitablely supplemented with glycine and pantothenate calcium. Conditioned medium (CM) stemming from cell suspension culture could remarkably promote the formation of single cell clone from culture cells of taxus below a critical cell plating density.