Institut Pasteur, Laboratoire de Technologie Cellulaire, 25, rue du Docteur Roux, F-75015, Paris.
Cytotechnology. 2000 Oct;34(1-2):175-9. doi: 10.1023/A:1008147822625.
The use of the ultrasonic separation deviceis a very important step in the direction forimproving animal cell bioreactor cultures. However,the normal construction of the ultrasonic separationdevice ADI 1015 has an inherent disadvantage inpumping the cell suspension continuously through thedevice by using a peristaltic pump. The cells aretaken out of the reactor and are transported to theside inlet located below the separation chamber of thedevice. This cycling leads to cell death and aconsiderable reduction of the viable cell density. Themodification of the configuration of the device (nocirculation of the cell suspension through theretention device; during approximately 9 minutescell-free supernatant is extracted; every 9 minute forabout one minute, the volume which is equivalent tothe interior volume of the chamber and the tubingconnecting the device to the reactor, is flushed backin order to return the retained cells back to thereactor) allows cell densities from 10(6) to2.7 x 10(6) c/ml with a viability of at least90% (tested for the shear sensitive insect cell lineHigh Five), whereas the maximal cell densitiesobtained were 0.76 x 10(6) c/ml for the periodof continuous culture and 10(5) c/ml at the end ofthe use of the device in the classical mode.
超声分离装置的使用是提高动物细胞生物反应器培养的一个非常重要的方向。然而,超声分离装置 ADI 1015 的正常结构在使用蠕动泵将细胞悬浮液连续通过装置泵送时存在固有缺点。细胞被从反应器中取出并输送到装置分离室下方的侧入口。这种循环导致细胞死亡和活细胞密度的显著降低。该装置结构的改进(细胞悬浮液不通过保留装置循环;大约 9 分钟内提取无细胞上清液;每 9 分钟大约 1 分钟,将相当于腔室和连接装置与反应器的管道的内部体积的体积冲洗回反应器,以将保留的细胞返回反应器)允许细胞密度从 10(6)到 2.7 x 10(6) c/ml,活率至少为 90%(对剪切敏感的昆虫细胞系 High Five 进行测试),而在连续培养期间获得的最高细胞密度为 0.76 x 10(6) c/ml,在经典模式下使用装置结束时为 10(5) c/ml。