Kompier R, Kislev N, Segal I, Kadouri A
Department of Biophysics, Weizmann Institute of Science, Rehovot, Israel.
Enzyme Microb Technol. 1991 Oct;13(10):822-7. doi: 10.1016/0141-0229(91)90066-j.
Insect cells (Spodoptera frugiperda) have been cultured in a stationary bed reactor, packed with a fibrous polyester carrier. When the bioreactor was perfused with serum-supplemented medium, a cell density of 6 x 10(6) cells ml-1 packed carrier was reached. Scanning electron microscopy investigations have shown that the insect cells grew along the three-dimensionally oriented fibers of the Fibra-cel carrier. After infection of the logarithmically growing cells with a recombinant baculovirus (Autographa californica) containing the gene coding for beta-galactosidase, the medium in the bioreactor was changed to serum-free medium. At day 13 postinfection (p.i.), a beta-galactosidase level of 320 microgram ml-1 and, at day 17 p.i., a virus titer of 2.1 x 10(8) TCID50 units ml-1 (day 17 p.i.) were reached. In another bioreactor, operated in a similar way but with serum-containing medium, a beta-galactosidase concentration of 360 microgram ml-1 and a virus titer of 2.3 x 10(8) TCID50 units ml-1 were obtained. These results indicate the potential use of this production system for the production of recombinant protein and baculovirus in insect cells.
昆虫细胞（草地贪夜蛾）已在装有纤维聚酯载体的固定床反应器中进行培养。当向生物反应器灌注补充了血清的培养基时，每毫升填充载体的细胞密度达到了6×10⁶个细胞。扫描电子显微镜研究表明，昆虫细胞沿着Fibra-cel载体的三维定向纤维生长。在用含有编码β-半乳糖苷酶基因的重组杆状病毒（苜蓿银纹夜蛾核型多角体病毒）感染对数生长期的细胞后，将生物反应器中的培养基更换为无血清培养基。在感染后第13天（p.i）β-半乳糖苷酶水平达到320微克/毫升，在感染后第17天（p.i）病毒滴度达到2.1×10⁸ TCID₅₀单位/毫升（感染后第17天）。在另一个以类似方式运行但使用含血清培养基操作的生物反应器中，获得了360微克/毫升的β-半乳糖苷酶浓度和2.3×10⁸ TCID₅₀单位/毫升的病毒滴度。这些结果表明该生产系统在昆虫细胞中生产重组蛋白和杆状病毒方面具有潜在用途。
