Cytotechnology

, Volume 56, Issue 1, pp 57–67 | Cite as

Separation of CHO cells using hydrocyclones

  • Rodrigo C. V. Pinto
  • Ricardo A. Medronho
  • Leda R. Castilho
Original Research

Abstract

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.

Keywords

Animal cells Cell separation CHO cells Hydrocyclones Perfusion Retention device 

Abbreviations

Dc

Hydrocyclone diameter (diameter of the cylindrical part)

Di

Inlet diameter

Do

Overflow diameter

Du

Underflow diameter

E

Separation efficiency

HC

Hydrocyclone

L

Hydrocyclone total length

\( {\ell } \)

Vortex finder length

Q

Feed flow rate

Qu

Underflow flow rate

Rf

Flow ratio

X

Cell concentration

Xu

Underflow cell concentration

ΔP

Pressure drop

ΔVu

Viability loss (cell viability in the feed minus viability in the underflow stream)

θ

Angle of the conical part

Notes

Acknowledgements

The authors wish to thank the Brazilian funding agencies CNPq, FAPERJ and FINEP for financial support.

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Copyright information

© Springer Science+Business Media B.V. 2007

Authors and Affiliations

  • Rodrigo C. V. Pinto
    • 1
  • Ricardo A. Medronho
    • 2
  • Leda R. Castilho
    • 1
  1. 1.Federal University of Rio de Janeiro (UFRJ), COPPE - Chemical Engineering ProgramRio de JaneiroBrazil
  2. 2.School of Chemistry, Chemical Engineering DepartmentFederal University of Rio de Janeiro (UFRJ)Rio de JaneiroBrazil

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