Vallez-Chetreanu F, Fraisse Ferreira L G, Rabe R, von Stockar U, Marison I W
Laboratory of Chemical and Biochemical Engineering, Ecole Polytechnique Fédérale de Lausanne (EPFL), 1015-Lausanne, Switzerland.
J Biotechnol. 2007 Jun 30;130(3):265-73. doi: 10.1016/j.jbiotec.2007.04.007. Epub 2007 Apr 24.
The main limitation in the use of spin-filters during perfusion cultures of animal cells was revealed to be filter fouling. This phenomenon involves cell-sieve interactions as well as cell attachment to, and growth on, the filter surface. The cell attachment effect has been analysed in the present study during long-term perfusion simulations with CHO animal cells. It was demonstrated that at low filter acceleration, below 6.2 m/s2, a high perfusion rate of 25 cm/h induced rapid filter pore clogging within 3 days, whereas increasing the filter acceleration to 25 m/s2 increased filter longevity from 3 to 25 days, for filters with a pore size of 8.5 microm. Increasing the filter pore size to 14.5 microm improved filter longevity by 84% with less viable and dead cell deposits on the filter surface. However, it was demonstrated that filter longevity was not necessarily dependent on the amount of cell deposit on the filter surface. In the second part of this study, ultrasonic technology was used to reduce filter fouling. Filter vibration, induced by a piezo actuator, improved filter longevity by 113% during CHO cells perfusion cultures.
动物细胞灌注培养过程中使用旋转过滤器的主要限制因素被发现是过滤器堵塞。这种现象涉及细胞与筛网的相互作用以及细胞在过滤器表面的附着和生长。在本研究中，使用CHO动物细胞进行长期灌注模拟时，对细胞附着效应进行了分析。结果表明，在低于6.2 m/s2的低过滤器加速度下，25 cm/h的高灌注速率会在3天内导致过滤器孔迅速堵塞，而对于孔径为8.5微米的过滤器，将过滤器加速度提高到25 m/s2可使过滤器寿命从3天延长至25天。将过滤器孔径增加到14.5微米可使过滤器寿命提高84%，且过滤器表面上存活和死亡细胞沉积物减少。然而，研究表明过滤器寿命不一定取决于过滤器表面上的细胞沉积物量。在本研究的第二部分，使用超声技术减少过滤器堵塞。在CHO细胞灌注培养过程中，由压电致动器引起的过滤器振动使过滤器寿命提高了113%。
