Departamento de Engenharia de Materiais e de Bioprocessos, Faculdade de Engenharia Química, Universidade Estadual de Campinas, Campinas, Brazil.
Biotechnol Adv. 2012 May-Jun;30(3):613-28. doi: 10.1016/j.biotechadv.2011.10.009. Epub 2011 Nov 4.
In the present review we discuss strategies that have been used for heterologous gene expression in Drosophila melanogaster Schneider 2 (S2) cells using plasmid vectors. Since the growth of S2 cells is not dependent on anchorage to solid substrates, these cells can be easily cultured in suspension in large volumes. The factors that most affect the growth and gene expression of S2 cells, namely cell line, cell passage, inoculum concentration, culture medium, temperature, dissolved oxygen concentration, pH, hydrodynamic forces and toxic metabolites, are discussed by comparison with other insect and mammalian cells. Gene expression, cell metabolism, culture medium formulation and parameters involved in cellular respiration are particularly emphasized. The experience of the authors with the successful expression of a biologically functional protein, the rabies virus glycoprotein (RVGP), by recombinant S2 cells is presented in the topics covered.
在本综述中，我们讨论了使用质粒载体在黑腹果蝇 Schneider 2（S2）细胞中进行异源基因表达的策略。由于 S2 细胞的生长不依赖于固体基质的附着，因此这些细胞可以很容易地在大量悬浮液中培养。通过与其他昆虫和哺乳动物细胞进行比较，讨论了影响 S2 细胞生长和基因表达的最重要因素，即细胞系、细胞传代、接种浓度、培养基、温度、溶解氧浓度、pH 值、水动力和有毒代谢物。特别强调了基因表达、细胞代谢、培养基配方和细胞呼吸涉及的参数。作者在涵盖的主题中介绍了通过重组 S2 细胞成功表达具有生物功能的蛋白质——狂犬病病毒糖蛋白（RVGP）的经验。
