Berson R Eric, Friederichs Goetz
Department of Chemical Engineering, University of Louisville, Louisville, Kentucky 40292, USA.
Biotechnol Prog. 2008 Jan-Feb;24(1):154-7. doi: 10.1021/bp070122w. Epub 2008 Jan 3.
The concept of a self-feeding roller bottle that delivers a continuous supply of fresh media to cells in culture, which is mechanically simplistic and works with existing roller apparatuses, is presented here. A conventional roller bottle is partitioned into two chambers; one chamber contains the fresh culture media reservoir, and the other contains the cell culture chamber. A spiroid of tubing inside the fresh media reservoir acts as a pump when the bottle rotates on its horizontal axis, continuously delivering fresh media through an opening in the partition to the cell culture chamber. The modified bottle proved capable of maintaining steady-state cell densities of a hybridoma cell line over the 10-day period tested, although at lower densities than reached during batch operation due to the continuous volume dilution. Steady-state density proved to be controllable by adjusting the perfusion rate, which changes with the rotation rate of the bottle. Specific antibody production rate is as much as 3.7 times the rate in conventional roller bottles operating with intermittent batch feeding.
本文介绍了一种自供料滚瓶的概念,它能为培养中的细胞持续供应新鲜培养基,其机械结构简单,可与现有的滚瓶装置配合使用。传统的滚瓶被分隔成两个腔室;一个腔室装有新鲜培养基储存器,另一个腔室装有细胞培养室。当瓶子绕其水平轴旋转时,新鲜培养基储存器内的螺旋形管道起到泵的作用,通过隔板上的开口将新鲜培养基持续输送到细胞培养室。经测试,改良后的瓶子在10天内能够维持杂交瘤细胞系的稳态细胞密度,不过由于持续的体积稀释,其密度低于分批操作时达到的密度。事实证明,通过调节灌注速率可以控制稳态密度,而灌注速率会随瓶子的旋转速度而变化。特异性抗体产生率比采用间歇分批进料操作的传统滚瓶高出3.7倍。