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題 名 | 中草藥資源永續使用之探討研究--建立枇杷細胞懸浮培養之研究=The Research for Traditional Herbal Resource Sustainable Utilization--Study on Cell Suspension Cultures of Eriobotrya japonica Lindl |
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作 者 | 吳金濱; | 書刊名 | 中醫藥年報 |
卷 期 | 光碟版1:10 2012.10[民101.10] |
頁 次 | 頁1-20 |
專 輯 | 中醫藥產業應用開發 |
分類號 | 418.5 |
關鍵詞 | 懸浮培養; 動力學研究; 二次代謝產物; 生物反應器; Suspension culture; Kinetic study; Secondary metabolites; Bioreactor; |
語 文 | 中文(Chinese) |
中文摘要 | 研究目的:本計畫主要探討枇杷細胞培養的過程中,養分消耗與二次代謝產物之間的動力學研究,進而探討兩者之間的交互作用與相互關係。在先前的研究中,已建立枇杷癒傷組織的理想培養環境。由於固體培養效率不高且不易分析,因此在本計畫進一步建立細胞懸浮震盪培養技術,進行養分與二次代謝產物之動力學研究。並且進一步建立 5L生物反應器培養技術。 研究方法:利用癒傷組織誘導懸浮細胞培養,經由馴化獲得高產率之細胞株,利用震盪懸浮培養建立各種巨量元素的分析方法。培養過程中,每兩天進行採樣,將取得的濾液進行各種養分含量的檢測;記錄細胞的生長情形與利用 HPLC分析細胞中二次代謝產物的含量。最後利用生物反應器試驗。 結果與討論:在 28天的震盪懸浮細胞培養中,蔗糖迅速分解為葡萄糖與果糖,第 18天消耗殆盡。磷酸鹽在細胞成長到第 10天時消耗殆盡;鎂與鈣離子在培養前期沒有明顯的變化,培養 18天後開始快速下降。此外,細胞重量的變化(4g/L→17g/L)方面,從第 10天開始重量的累積加快;在第 18天之後,細胞的重量停止增加,進入細胞生長老化期。有用二次代謝產物的累積從 18天後開始快速增加。根據本實驗結果,當導電度低於 4μm/cm,或是碳源消耗殆盡時,細胞開始快速的消耗培養液中的鈣離子與鎂離子,細胞重量停止增加並且迅速的累積二次代謝產物含量。而且在生物反應器的初步研究中,結果顯示利用生物反應器培養更縮短生長週期(16天)、產生更多的細胞(25g/L)與二次代謝產物(160mg/g)。 |
英文摘要 | Aim: The aim of this study is to investigate the relation in nutrient consumption and secondary metabolites kinetics on the suspension culture of Eriobotryae cells. On the previous study, we had established the optimal culture state in loquat tissue culture. However, the solid culture has low efficiency and hard to analysis. In the study, we established suspension culture of Eriobotryae cell and analysis the kinetic factors between cell growth and metabolism. Also, the 5L bioreactor culture was established. Method: To induce the suspension cells and high productivity cell line, and establish the analysis methods of scale inorganic elements. The harvesting of cells and collection of spent medium were carried out with control cultures every 2 d until 24 d. The cell weight was recorded, the determination of residual sugar and ions in the medium was carried out, and analysis the secondary metabolites contain. Results & Discussion: In 28 days cultivation, the result showed that the sucrose rapidly decomposed to glucose and fructose, and totally consumed on day 18. The phosphate was totally consumed on day 10, while the magnesium and calcium revealed no evident changes until day 18. The cell weight accumulation increased on day 10, and stopped on day 18, while the secondary metabolites started to accumulate rapidly on day 18. According to the result and data previously, when the conductivity is under 4μm/cm, or the carbon source is empty, the cell will uptake the magnesium and calcium from the medium, while the cell weight stop increasing and start to accumulate the secondary metabolites. In the bioreactor experiment, the result showed that the bioreactor culture has a shorter culture period, better cell growth and better secondary metabolites accumulation. |
本系統中英文摘要資訊取自各篇刊載內容。