Swiech Kamilla, da Silva Clóvis S, Arantes Mabel K, dos Santos Alexandra S, Astray Renato M, Pereira Carlos A, Suazo Cláudio A T
Departamento de Engenharia Química, Universidade Federal de São Carlos, Via Washington Luis, Km 235, CEP 13565-905, São Carlos-SP, Brazil.
Biotechnol Appl Biochem. 2008 Jan;49(Pt 1):41-9. doi: 10.1042/BA20060148.
In the present study, the growth and key metabolic features of a gene-transfected Drosophila melanogaster (fruitfly) S2 (Schneider 2) cell population (S2AcRVGP cells), cultured in Sf900-II medium, have been evaluated to provide substantial support for the development of a bioprocess to produce RVGP (rabies-virus glycoprotein). Experimental cultures were grown both in a 100 ml Schott flask incubated in a shaker at 28 degrees C and 100 rev./min and in a 3 litre stirred-tank bioreactor at 28 degrees C, with increasing agitation. In small-scale culture, S2AcRVGP cells reached a maximum cell concentration of 1.13 x 10(7) cell/ml, presented a mu(max) (maximum specific growth rate) of 0.037 h(-1) and the growth was limited by oxygen deprivation. An early and remarkably long stationary phase was observed under hypoxia. Cell cultures grown in the bioreactor without oxygen limitation exhibited a maximum cell concentration of 2.2 x 10(7) cells/ml and mu(max) values as high as 0.048 h(-1). The main substrate consumed in order to reach such a high growth rate was the amino acid proline, which seems to play an important role as a source of metabolic energy in the culture of S2AcRVGP cells. Under conditions of hypoxia, the cells were able to survive for 15 h without apparent damage, recovering their previous metabolic activity.
在本研究中，对在Sf900-II培养基中培养的基因转染的黑腹果蝇（果蝇）S2（施奈德2）细胞群体（S2AcRVGP细胞）的生长和关键代谢特征进行了评估，为开发生产狂犬病病毒糖蛋白（RVGP）的生物工艺提供了重要支持。实验培养在28℃、转速为100转/分钟的摇床上孵育的100毫升肖特烧瓶中进行，也在28℃的3升搅拌罐生物反应器中进行，搅拌速度逐渐增加。在小规模培养中，S2AcRVGP细胞达到的最大细胞浓度为1.13×10⁷个细胞/毫升，最大比生长速率（μmax）为0.037 h⁻¹，生长受到缺氧限制。在缺氧条件下观察到早期且显著较长的稳定期。在无氧气限制的生物反应器中培养的细胞培养物表现出的最大细胞浓度为2.2×10⁷个细胞/毫升，μmax值高达0.048 h⁻¹。为达到如此高的生长速率而消耗的主要底物是氨基酸脯氨酸，脯氨酸似乎在S2AcRVGP细胞培养中作为代谢能量来源发挥重要作用。在缺氧条件下，细胞能够在无明显损伤的情况下存活15小时，并恢复其先前的代谢活性。
