Escola Politécnica da Usp - Departamento de Engenharia Química , Av. Prof. Luciano Gualberto, trav.3 n.380 - PQI/EPUSP , São Paulo, SP, 05508-970, Brazil,
Cytotechnology. 2008 May;57(1):37-44. doi: 10.1007/s10616-007-9118-8. Epub 2008 Feb 27.
Specific respiration rate ( [Formula: see text]) is a key parameter to understand cell metabolism and physiological state, providing useful information for process supervision and control. In this work, we cultivated different insect cells in a very controlled environment, being able to measure [Formula: see text]. Spodoptera frugiperda (Sf9) cells have been used through virus infection as host for foreign protein expression and bioinsecticide production. Transfected Drosophila melanogaster (S2) cells can be used to produce different proteins. The objective of this work is to investigate respiratory activity and oxygen transfer during the growth of different insect cells lines as Spodoptera frugiperda (Sf9), Drosophila melanogaster (S2) wild and transfected for the expression of GPV and EGFP. All experiments were performed in a well-controlled 1-L bioreactor, with SF900II serum free medium. Spodoptera frugiperda (Sf9) cells reached 10.7 x 10(6) cells/mL and maximum specific respiration rate ([Formula: see text]) of 7.3 x 10(-17) molO(2)/cell s. Drosophila melanogaster (S2) cells achieved 51.2 x 10(6) cells/mL and [Formula: see text] of 3.1 x 10(-18) molO(2)/cell s. S2AcGPV (expressing with rabies virus glycoprotein) reached 24.9 x 10(6) cells/mL and [Formula: see text] of 1.7 x 10(-17) molO(2)/cell s, while S2MtEGFP (expressing green fluorescent protein) achieved 15.5 x 10(6) cells/mL and [Formula: see text] = 1.9 x 10(-17) molO(2)/cell s. Relating to the Sf9, S2 cells reached higher maximum cell concentrations and lower specific respiration rate, which can be explained by its smaller size. These results presented useful information for scale-up and process control of insect cells.
特定呼吸率([Formula: see text])是理解细胞代谢和生理状态的关键参数,为过程监测和控制提供了有用的信息。在这项工作中,我们在非常受控的环境中培养了不同的昆虫细胞,能够测量[Formula: see text]。草地贪夜蛾(Sf9)细胞已被用于病毒感染作为宿主进行外源蛋白表达和生物杀虫剂生产。转染的黑腹果蝇(S2)细胞可用于生产不同的蛋白质。这项工作的目的是研究不同昆虫细胞系的呼吸活性和氧气传递,包括草地贪夜蛾(Sf9)、黑腹果蝇(S2)野生型和转染用于表达 GPV 和 EGFP 的细胞。所有实验均在一个控制良好的 1 升生物反应器中进行,使用 SF900II 无血清培养基。草地贪夜蛾(Sf9)细胞达到 10.7 x 10(6)个细胞/mL,最大特定呼吸率([Formula: see text])为 7.3 x 10(-17) molO(2)/细胞 s。黑腹果蝇(S2)细胞达到 51.2 x 10(6)个细胞/mL,[Formula: see text]为 3.1 x 10(-18) molO(2)/细胞 s。S2AcGPV(表达狂犬病病毒糖蛋白)达到 24.9 x 10(6)个细胞/mL,[Formula: see text]为 1.7 x 10(-17) molO(2)/细胞 s,而 S2MtEGFP(表达绿色荧光蛋白)达到 15.5 x 10(6)个细胞/mL,[Formula: see text]为 1.9 x 10(-17) molO(2)/细胞 s。与 Sf9 相比,S2 细胞达到了更高的最大细胞浓度和更低的特定呼吸率,这可以用其较小的尺寸来解释。这些结果为昆虫细胞的放大和过程控制提供了有用的信息。