台灣紅豆杉之細胞培養與紫杉烷類生產

比較五個不同來源之台灣紅豆杉(Taxus mairei)細胞系的生長與紫杉烷類產量，發現細胞系間差異很大；細胞系A生長最好，但完全不含任何taxane，細胞系D生長中等，紫杉醇含量為16.8mg/L，是為用於生產紫杉醇之最佳細胞系。以細胞系D進行細胞培養試驗，發現細胞培養開始的接種密度與舊培養液之保留會影響細胞的生長。在鮮重100g/L之細胞密度下進行培養，細胞生長最佳，細胞加倍的時間為6d，培養24d後不論是體積、乾重或鮮重都可增加3-3.5倍。繼代培養時以含細胞之舊培養液與新鮮培養基比例為3:7與4:6時細胞生長最好；若只取少量或完全不加舊培養基會使培養基與細胞變紅色，且生長受阻。以1L三角瓶建立細胞連續培養方法，每15d更換部分培養基之方式培養，可使細胞在低密度(25-50g/L)接種條件下，在60d後細胞體積增加10.7-11.5倍。

關鍵字

台灣紅豆杉；細胞培養；紫杉烷類化合物；紫杉醇

並列摘要

Cell cultures derived from 5 different sources of Taxtus mairei were established. Both cell growth and taxane contents varied greatly among the 5 cell lines. Cell line A grew the most rapid but contained no taxane, while cell line D grew moderately but contained 16.8 mg/L of taxol, greater than that of the other cell lines. In vitro cultures of cell line D were used to establish cell culture methods. We found that cell growth in flasks was affected as cell densities changed and we used a medium volume in initial cultures. The optimal initial cell density was 100 g/L which gave a cell volume that doubled by 6 d. with 3- to 3.5-fold increments in terms of cell volume, and fresh and dry weights after 24 d in culture. The optimal ratios of used to fresh medium utilized for the growth of subcultures were 3:7 and 4:6. Reducing the volume of used medium in subcultures caused cells to turn red and be stunted. Continuous cultures in 1-L flasks were established by renewing the medium once every 15 d in a 60-d culturing period. Although the initial cell densities were 25 to 50 g/L, lower than those recorded in previous tests, the final cell volumes increased 10.7-11.5 times, respectively.

並列關鍵字

Taxtus mairei ； cell culture ； taxane ； taxol

國際替代計量

台灣紅豆杉之細胞培養與紫杉烷類生產

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