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

Research output: Contribution to journalArticlepeer-review


  • Mariana P. Hanga
  • Karen Coopman
  • Christopher J. Hewitt

Colleges, School and Institutes

External organisations

  • Loughborough University


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
Number of pages13
JournalJournal of Chemical Technology and Biotechnology
Issue number7
Early online date14 Mar 2017
Publication statusPublished - 7 Jun 2017


  • expansion, human bone marrow-derived mesenchymal stem/stromal cells (hMSCs), liquid/liquid interface, perfluorocarbons, two-phase system