The Use of Hydrocyclones for Mammalian Cell Retention in Perfusion Bioreactors

Jockwer, Alexander; Medronho, Ricardo de Andrade; Wagner, Roland; Anspach, Friedrich Birger; Deckwer, Wolf‐Dieter

doi:10.1007/978-94-010-0369-8_69

Cited by 22 publications

(23 citation statements)

References 4 publications

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“…These results suggest that there may be a difference between HeLa and CHO cells regarding sensitivity to shear generated inside hydrocyclones. Furthermore, comparing the present results with those obtained for CHO cells grown in serum-free medium (Jockwer et al 2001), the data also suggest that the presence of seric proteins could be attenuating the effects of the hydrodynamic stress on cells maintained in serumsupplemented medium, as previously observed by Van Der Pol et al (1990).…”

Section: Resultssupporting

confidence: 89%

“…Furthermore, when compared to other hydrocyclone geometries, the efficiencies found in the present work are higher than the best efficiency (81%) obtained for HeLa cells by Lübberstedt et al (2000b). These authors used commercial hydrocyclones designed for conventional (non-biological) The viability losses found in the present work were slightly higher than those obtained with commercial Dorr-Oliver hydrocyclones for HeLa cells (Lübber-stedt et al 2000a), but lower than those obtained by Jockwer et al (2001) for the separation of CHO cells cultured in serum-free medium, using the same type of hydrocyclone as in this work. Just in the case of HC 2020, viability loss was identical (14.4%) in this study and in that by Jockwer et al (2001).…”

Section: Resultscontrasting

confidence: 52%

“…These authors used commercial hydrocyclones designed for conventional (non-biological) The viability losses found in the present work were slightly higher than those obtained with commercial Dorr-Oliver hydrocyclones for HeLa cells (Lübber-stedt et al 2000a), but lower than those obtained by Jockwer et al (2001) for the separation of CHO cells cultured in serum-free medium, using the same type of hydrocyclone as in this work. Just in the case of HC 2020, viability loss was identical (14.4%) in this study and in that by Jockwer et al (2001). These results suggest that there may be a difference between HeLa and CHO cells regarding sensitivity to shear generated inside hydrocyclones.…”

Section: Resultscontrasting

confidence: 43%

“…Hydrocyclones have been proposed in the recent years to promote animal cell separation in batch and perfusion processes (Lübberstedt et al 2000a, b;Jockwer et al 2001). These separation devices present several advantages for use in the biotechnology industry, such as their simplicity and reliability, as well as the absence of moving parts.…”

Section: Introductionmentioning

confidence: 99%

“…Although the shear stress levels inside hydrocyclones are relatively high, the residence times of the cells inside the equipment are very short (in the range of fractions of seconds), specially for those cells being separated in the underflow concentrated cell stream, which is the stream that is recycled to the bioreactor in a perfusion culture. Jockwer et al (2001) studied hydrocyclones specially designed for animal cell separation and were able to successfully carry out a 23-day perfusion culture in a 5-L bioreactor with CHO (Chinese hamster ovary) cells.…”

Section: Introductionmentioning

confidence: 99%

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Separation of CHO cells using hydrocyclones

2007

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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 wildtype 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.

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Section: Resultssupporting

confidence: 89%