Expansion of bone marrow-derived human mesenchymal stem/stromal cells (hMSCs) using a two-phase liquid/liquid system

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

*Corresponding author for this work

Research output: Contribution to journalArticle

Abstract

BACKGROUND: Human mesenchymal stem/stromal cells (hMSCs) are at the forefront of regenerative medicine applications due to their relatively easy isolation and availability in adults, potential to differentiate and to secrete a range of trophic factors that could determine specialised tissue regeneration. To date, hMSCs have been successfully cultured in vitro on substrates such as polystyrene dishes (TCPS) or microcarriers. However, hMSC sub-cultivation and harvest typically employs proteolytic enzymes that act by cleaving important cell membrane proteins resulting in long-term cell damage. In a process where the cells themselves are the product, a non-enzymatic and non-damaging harvesting approach is desirable.
RESULTS: An alternative system for hMSC expansion and subsequent non-enzymatic harvest was investigated here. A liquid/liquid two-phase system was proposed, comprising a selected perfluorocarbon (FC40) and growth medium (DMEM). The cells exhibited similar cell morphologies compared with TCPS. Moreover, they retained their identity and differentiation potential post-expansion and post-harvest. Further, no significant difference was found when culturing hMSCs in the culture systems prepared with either fresh or recycled FC40 perfluorocarbon.
CONCLUSIONS: These findings make the FC40/DMEM system an attractive alternative for traditional cell culture substrates due to their ease of cell recovery and recyclability, the latter impacting on overall process costs.
Original languageEnglish
Pages (from-to)1577-1589
JournalJournal of Chemical Technology and Biotechnology
Volume92
Issue number7
Early online date14 Mar 2017
DOIs
Publication statusPublished - 7 Jun 2017

Fingerprint

Stem cells
Mesenchymal Stromal Cells
bone
Bone
Bone Marrow
stem
liquid
Liquids
Fluorocarbons
Cell culture
substrate
Tissue regeneration
Polystyrenes
Substrates
Cell membranes
medicine
Membrane Proteins
Peptide Hydrolases
Cell Culture Techniques
regeneration

Bibliographical note

© 2017 The Authors. Journal of Chemical Technology & Biotechnology published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry. This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.

Funding: BBSRC (BRIC BB/K011066/1 and BB/K01099/1).

Keywords

  • human mesenchymal stem cells (hMSCs)
  • two-phase system
  • expansion
  • liquid/liquid interface
  • perfluorocarbons

Cite this

Hanga, Mariana ; Murasiewicz, Halina ; Pacek, Andrzej W. ; Nienow, Alvin W. ; Coopman, Karen ; Hewitt, Christopher. / Expansion of bone marrow-derived human mesenchymal stem/stromal cells (hMSCs) using a two-phase liquid/liquid system. In: Journal of Chemical Technology and Biotechnology. 2017 ; Vol. 92, No. 7. pp. 1577-1589.
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Expansion of bone marrow-derived human mesenchymal stem/stromal cells (hMSCs) using a two-phase liquid/liquid system. / Hanga, Mariana; Murasiewicz, Halina; Pacek, Andrzej W.; Nienow, Alvin W.; Coopman, Karen; Hewitt, Christopher.

In: Journal of Chemical Technology and Biotechnology, Vol. 92, No. 7, 07.06.2017, p. 1577-1589.

Research output: Contribution to journalArticle

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T1 - Expansion of bone marrow-derived human mesenchymal stem/stromal cells (hMSCs) using a two-phase liquid/liquid system

AU - Hanga, Mariana

AU - Murasiewicz, Halina

AU - Pacek, Andrzej W.

AU - Nienow, Alvin W.

AU - Coopman, Karen

AU - Hewitt, Christopher

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PY - 2017/6/7

Y1 - 2017/6/7

N2 - BACKGROUND: Human mesenchymal stem/stromal cells (hMSCs) are at the forefront of regenerative medicine applications due to their relatively easy isolation and availability in adults, potential to differentiate and to secrete a range of trophic factors that could determine specialised tissue regeneration. To date, hMSCs have been successfully cultured in vitro on substrates such as polystyrene dishes (TCPS) or microcarriers. However, hMSC sub-cultivation and harvest typically employs proteolytic enzymes that act by cleaving important cell membrane proteins resulting in long-term cell damage. In a process where the cells themselves are the product, a non-enzymatic and non-damaging harvesting approach is desirable. RESULTS: An alternative system for hMSC expansion and subsequent non-enzymatic harvest was investigated here. A liquid/liquid two-phase system was proposed, comprising a selected perfluorocarbon (FC40) and growth medium (DMEM). The cells exhibited similar cell morphologies compared with TCPS. Moreover, they retained their identity and differentiation potential post-expansion and post-harvest. Further, no significant difference was found when culturing hMSCs in the culture systems prepared with either fresh or recycled FC40 perfluorocarbon. CONCLUSIONS: These findings make the FC40/DMEM system an attractive alternative for traditional cell culture substrates due to their ease of cell recovery and recyclability, the latter impacting on overall process costs.

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