攪拌式生物反應器容器縐折設計對溶氧傳遞及混合效能之影響與動態測定法估算香蕉胚性細胞生長

Oxygen Transfer and Mixing Performance in Designed Crease Bioreactor, and Using Dynamic Technique to Estimate the Embryogenic Cell Growth of Banana

體積為3L之圓筒形玻璃容器，其構形經縐折改變後，利用旋轉攪拌方式設計做為植物細胞培養之生物反應器。當通入低流量的氮氣逐氧或導入低流量的空氣，顯示經縐折構形改變之容器具有較佳之保氧與溶氧能力。以亞硫酸鈉法計算容量氧氣傳遞係數（KLa）值，在低流量通氣0.16 vvm,5 psi之攪拌下，隨攪拌旋轉速度增加，KLa亦增加。縐折設計與不經改變之反應槽比較其KLa，攪拌速度達110 rpm時，兩者依序平均為8.59與5.67（hr^(-1)）。由流體染色微珠追蹤其旋轉一週便可上下液面一次，而不經改變之反應器約需旋轉四週才達液面。以香蕉細胞接種在縐折設計之生物反應器，胚性細胞培養環境控制在20.0±5.0%溶氧濃度，較佳之通氣方式為低流量並間歇式空氣導入方式，旋轉攪拌以110 rpm，除可避免植物細胞剪切力敏感影響，亦可減低氣泡堆積現象。監控溶氧變化之動態測定法，可提出植物細胞合理之生長量曲線，並便於規模化生產操作，取代細胞沉積體積因植物細胞聚集化現象，所造成體積沉積之誤差。 The 3L volume of spinner bioreactor was modified to the creased conformation. The dynamic gasout technique was operated through lower flow rate of nitrogen and air, the creased vessel showed high capability of retaining and dissolved oxygen in water. Comparison between creased and round vessel, the volumetric oxygen transfer coefficient (K(subscript L) a) was estimated 8.59 and 5.67 (hr^(-1)) by sodium sulfite technique under aeration 0.16 vvm, 5 psi of air and 110 rpm agitation rate in water. Increased agitation rate significantly enhanced the value of K(subscript L) a. Using minibead tracers, one fluid pathway of crease vessel could approach the gas interface once. In contrast, it took four circles for the round vessel to reach the interface. In embryogenic cells of triploid banana cultured in designed crease bioreactor, the better option was to control dissolved oxygen at 20.0±5.0%, 110 rpm agitated rate and aerated with intermittence. The designed conditions could protect plant cells against the shear force, and reduce the bleb accumulation upon the face of liquid. Dynamic technique monitoring dissolved oxygen concentration could better estimate cell growth curve than cell packed volume method. Current design of creased bioreactor is idea for scaleup plant cell culture.