Francis K M, O'Connor K C, Spaulding G F
Department of Chemical Engineering, Tulane University, New Orleans, Louisiana 70118, USA.
In Vitro Cell Dev Biol Anim. 1997 May;33(5):332-6. doi: 10.1007/s11626-997-0002-9.
A methodology is presented to culture Fall Armyworm Ovary cells in simulated micrograviy using a novel bioreactor developed by NASA, the High-Aspect Ratio Vessel. In this vessel, the growth and metabolic profile for these insect cells were profoundly different than those obtained in shaker-flask culture. Specifically, stationary phase in the NASA vessel was extended from 24 h to at least 7 d while cell concentration and viability remained in excess of 1 x 10(7) viable cells/ml and 90%, respectively. Measurements of glucose utilization, lactate production, ammonia production, and pH change indicate that simulated microgravity had a twofold effect on cell metabolism. Fewer nutrients were consumed and fewer wastes were produced in stationary phase by as much as a factor of 4 over that achieved in shaker culture. Those nutrients that were consumed in the NASA vessel were directed along different metabolic pathways as evidenced by an extreme shift in glucose utilization from consumption to production in lag phase and a decrease in yield coefficients by one half in stationary phase. These changes reflect a reduction in hydrodynamic forces from over 1 dyne/cm2 in shaker culture to under 0.5 dyne/cm2 in the NASA vessel. These results suggest that cultivation of insect cells in simulated microgravity may reduce production costs of cell-derived biologicals by extending production time and reducing medium requirements.
本文介绍了一种利用美国国家航空航天局（NASA）开发的新型生物反应器——高纵横比容器，在模拟微重力条件下培养草地贪夜蛾卵巢细胞的方法。在该容器中，这些昆虫细胞的生长和代谢特征与摇瓶培养中获得的显著不同。具体而言，NASA容器中的稳定期从24小时延长至至少7天，而细胞浓度和活力分别保持在超过1×10⁷个活细胞/毫升和90%以上。对葡萄糖利用、乳酸产生、氨产生和pH变化的测量表明，模拟微重力对细胞代谢有双重影响。在稳定期，与摇瓶培养相比，消耗的营养物质减少，产生的废物减少多达4倍。在NASA容器中消耗的那些营养物质沿着不同的代谢途径进行，这在滞后期葡萄糖利用从消耗到产生的极端转变以及稳定期产率系数降低一半中得到证明。这些变化反映了流体动力从摇瓶培养中的超过1达因/平方厘米降低到NASA容器中的低于0.5达因/平方厘米。这些结果表明，在模拟微重力条件下培养昆虫细胞可能通过延长生产时间和减少培养基需求来降低细胞衍生生物制品的生产成本。
