Federal University of Rio de Janeiro (UFRJ), COPPE - Chemical Engineering Program, Caixa Postal 68502, CEP 21941-972, Rio de Janeiro, RJ, Brazil.
Cytotechnology. 2008 Jan;56(1):57-67. doi: 10.1007/s10616-007-9108-x. Epub 2007 Nov 14.
Hydrocyclones are simple and robust separation devices with no moving parts. In the past few years, their use in animal cell separation has been proposed. In this work, the use of different hydrocyclone configurations for Chinese hamster ovary (CHO) cell separation was investigated following an experimental design. It was shown that cell separation efficiencies for cultures of the wild-type CHO.K1 cell line and of a recombinant CHO cell line producing granulocyte-macrophage colony stimulating factor (GM-CSF) were kept above 97%. Low viability losses were observed, as measured by trypan blue exclusion and by determination of intracellular lactate dehydrogenase (LDH) released to the culture medium. Mathematical models were proposed to predict the flow rate, flow ratio and separation efficiency as a function of hydrocyclone geometry and pressure drop. When cells were monitored for any induction of apoptosis upon passage through the hydrocyclones, no increase in apoptotic cell concentration was observed within 48 h of hydrocycloning. Thus, based on the high separation efficiencies, the robustness of the equipment, and the absence of apoptosis induction, hydrocyclones seem to be specially suited for use as cell retention devices in long-term perfusion runs.
水力旋流器是一种简单而坚固的分离设备,没有运动部件。在过去的几年中,已经有人提议将其用于动物细胞分离。在这项工作中,采用实验设计研究了不同水力旋流器结构在仓鼠卵巢(CHO)细胞分离中的应用。结果表明,对于野生型 CHO.K1 细胞系和产生粒细胞-巨噬细胞集落刺激因子(GM-CSF)的重组 CHO 细胞系的培养物,细胞分离效率均保持在 97%以上。通过台盼蓝排斥试验和测定细胞内乳酸脱氢酶(LDH)释放到培养液中的方法,观察到细胞活力损失较低。提出了数学模型来预测流量、流量比和分离效率作为水力旋流器几何形状和压降的函数。当通过水力旋流器监测细胞是否发生凋亡诱导时,在水力旋流 48 小时内,未观察到凋亡细胞浓度增加。因此,基于高分离效率、设备的坚固性和无凋亡诱导,水力旋流器似乎特别适合用作长期灌注运行中的细胞保留装置。