Engineering considerations on the use of liquid/liquid two-phase systems as a cell culture platform

Halina Murasiewicz*, Alvin W. Nienow, Mariana P. Hanga, Karen Coopman, Christopher J. Hewitt, Andrzej W. Pacek

*Corresponding author for this work

Research output: Contribution to journalArticle

Abstract

BACKGROUND: Application of perfluorocarbon based liquid/liquid two-phase systems for cell culture expansion has been investigated at small scale for more than 30 years and it has been established that such systems are able to support the survival of a variety of cell lines. Application of drops in liquid/liquid dispersions as temporary microcarriers is an exciting prospect as it enables adherent cells to be grown in stirred bioreactors, without the need to use enzymatic dissociation methods to harvest the cells. RESULTS: Two aspects of scaling up of perfluorocarbon/cell culture medium dispersions were investigated: (i) the effect of processing conditions on drop size/interfacial area; and (ii) the kinetics of separation of a stagnant dispersion. The processing conditions to produce the stable 'liquid microcarriers' with average drop size between 150 and 220μm have been established. Separation of dispersion into two continuous systems requires complete removal of proteins from the perfluorocarbon/cell culture medium interface. CONCLUSIONS: The correlation relating average drop size to the energy input and physical properties of both phases was developed and the method of separation of stable perfluorocarbon/cell culture medium dispersion was established. As the perfluorocarbon does not deteriorate during cell expansion and subsequent separation followed by sterilization, it could be re-used, making application of such systems at large scale very attractive and economical.

Original languageEnglish
Pages (from-to)1690-1698
JournalJournal of Chemical Technology and Biotechnology
Volume92
Issue number7
Early online date6 Feb 2017
DOIs
Publication statusPublished - 7 Jun 2017

Fingerprint

Fluorocarbons
Cell culture
Cell Culture Techniques
engineering
liquid
Liquids
Culture Media
Dispersions
Bioreactors
Processing
bioreactor
Physical properties
physical property
Cells
Proteins
Cell Line
kinetics
Kinetics
protein
energy

Bibliographical note

This is the peer reviewed version of the following article: Murasiewicz, H., Nienow, A. W., Hanga, M. P., Coopman, K., Hewitt, C. J., & Pacek, A. W. (2017). Engineering considerations on the use of liquid/liquid two-phase systems as a cell culture platform. Journal of Chemical Technology and Biotechnology, Early view. , which has been published in final form at http://dx.doi.org/10.1002/jctb.5166. This article may be used for non-commercial purposes in accordance with Wiley Terms and Conditions for Self-Archiving.

Funding: BBSRC BRIC. Grant Numbers: BB/K011066/1, BB/K01099/1

Keywords

  • bioreactors
  • cell culture
  • emulsification/dispersion
  • liquid microcarriers
  • perfluorocarbon

Cite this

Murasiewicz, Halina ; Nienow, Alvin W. ; Hanga, Mariana P. ; Coopman, Karen ; Hewitt, Christopher J. ; Pacek, Andrzej W. / Engineering considerations on the use of liquid/liquid two-phase systems as a cell culture platform. In: Journal of Chemical Technology and Biotechnology. 2017 ; Vol. 92, No. 7. pp. 1690-1698.
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Engineering considerations on the use of liquid/liquid two-phase systems as a cell culture platform. / Murasiewicz, Halina; Nienow, Alvin W.; Hanga, Mariana P.; Coopman, Karen; Hewitt, Christopher J.; Pacek, Andrzej W.

In: Journal of Chemical Technology and Biotechnology, Vol. 92, No. 7, 07.06.2017, p. 1690-1698.

Research output: Contribution to journalArticle

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T1 - Engineering considerations on the use of liquid/liquid two-phase systems as a cell culture platform

AU - Murasiewicz, Halina

AU - Nienow, Alvin W.

AU - Hanga, Mariana P.

AU - Coopman, Karen

AU - Hewitt, Christopher J.

AU - Pacek, Andrzej W.

N1 - This is the peer reviewed version of the following article: Murasiewicz, H., Nienow, A. W., Hanga, M. P., Coopman, K., Hewitt, C. J., & Pacek, A. W. (2017). Engineering considerations on the use of liquid/liquid two-phase systems as a cell culture platform. Journal of Chemical Technology and Biotechnology, Early view. , which has been published in final form at http://dx.doi.org/10.1002/jctb.5166. This article may be used for non-commercial purposes in accordance with Wiley Terms and Conditions for Self-Archiving. Funding: BBSRC BRIC. Grant Numbers: BB/K011066/1, BB/K01099/1

PY - 2017/6/7

Y1 - 2017/6/7

N2 - BACKGROUND: Application of perfluorocarbon based liquid/liquid two-phase systems for cell culture expansion has been investigated at small scale for more than 30 years and it has been established that such systems are able to support the survival of a variety of cell lines. Application of drops in liquid/liquid dispersions as temporary microcarriers is an exciting prospect as it enables adherent cells to be grown in stirred bioreactors, without the need to use enzymatic dissociation methods to harvest the cells. RESULTS: Two aspects of scaling up of perfluorocarbon/cell culture medium dispersions were investigated: (i) the effect of processing conditions on drop size/interfacial area; and (ii) the kinetics of separation of a stagnant dispersion. The processing conditions to produce the stable 'liquid microcarriers' with average drop size between 150 and 220μm have been established. Separation of dispersion into two continuous systems requires complete removal of proteins from the perfluorocarbon/cell culture medium interface. CONCLUSIONS: The correlation relating average drop size to the energy input and physical properties of both phases was developed and the method of separation of stable perfluorocarbon/cell culture medium dispersion was established. As the perfluorocarbon does not deteriorate during cell expansion and subsequent separation followed by sterilization, it could be re-used, making application of such systems at large scale very attractive and economical.

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