Deutschmann S M, Jäger V
Gesellschaft für Biotechnologische Forschung mbH, Arbeitsgruppe Zellkulturtechnik, Braunschweig, Germany.
Enzyme Microb Technol. 1994 Jun;16(6):506-12. doi: 10.1016/0141-0229(94)90022-1.
Spodoptera frugiperda insect cells (IPLB-Sf21-AE) (Sf21), infected with baculovirus expression vectors during their exponential growth phase, are commonly used to produce a variety of heterologous recombinant proteins. In the present study the culture conditions of these insect cells were studied to establish high-density suspension cultures with prolonged exponential growth phases. The Sf21 cells were grown in 125-ml spinner flasks using five different culture media supplemented with 5% fetal calf serum and four protein-free or low-protein culture media. The best results were achieved in EX-CELL 401 (protein-free media) and in IPL-41 modified with 2.5 g l-1 tryptose phosphate broth (serum-supplemented media), respectively. The latter was used for further batch and continuous cultivation of Sf21 cells in a perfused 1.4-l stirred-tank bioreactor with special attention to the oxygen requirement of these cells. Optimal growth was found at an oxygen concentration of 70% air saturation, resulting in a prolonged exponential growth phase that could be maintained for more than 16 days. A maximum cell density of 5.5 x 10(7) viable cells ml-1 was achieved.
草地贪夜蛾昆虫细胞（IPLB-Sf21-AE）（Sf21）在指数生长期感染杆状病毒表达载体，常用于生产多种异源重组蛋白。在本研究中，对这些昆虫细胞的培养条件进行了研究，以建立具有延长指数生长期的高密度悬浮培养。Sf21细胞在125毫升转瓶中培养，使用五种添加5%胎牛血清的不同培养基以及四种无蛋白或低蛋白培养基。分别在EX-CELL 401（无蛋白培养基）和添加2.5克/升磷酸胰蛋白胨肉汤的IPL-41（添加血清培养基）中获得了最佳结果。后者用于在1.4升灌注搅拌罐生物反应器中对Sf21细胞进行进一步的分批和连续培养，特别关注这些细胞的氧气需求。在氧气浓度为70%空气饱和度时发现了最佳生长，从而产生了可维持超过16天的延长指数生长期。实现了5.5×10⁷个活细胞/毫升的最大细胞密度。
