Exploring continuous and integrated strategies for the up- and downstream processing of human mesenchymal stem cells

Bárbara Cunha, Tiago Aguiar, Marta M. Silva, Ricardo J S Silva, Marcos F Q Sousa, Earl Pineda, Cristina Peixoto, Manuel José Teixeira Carrondo, Margarida Serra, Paula M. Alves

Research output: Contribution to journalArticle

27 Citations (Scopus)

Abstract

The integration of up- and downstream unit operations can result in the elimination of hold steps, thus decreasing the footprint, and ultimately can create robust closed system operations. This type of design is desirable for the bioprocess of human mesenchymal stem cells (hMSC), where high numbers of pure cells, at low volumes, need to be delivered for therapy applications. This study reports a proof of concept of the integration of a continuous perfusion culture in bioreactors with a tangential flow filtration (TFF) system for the concentration and washing of hMSC. Moreover, we have also explored a continuous alternative for concentrating hMSC.Results show that expanding cells in a continuous perfusion operation mode provided a higher expansion ratio, and led to a shift in cells' metabolism. TFF operated either in continuous or discontinuous allowed to concentrate cells, with high cell recovery (>80%) and viability (>95%); furthermore, continuous TFF permitted to operate longer with higher cell concentrations. Continuous diafiltration led to higher protein clearance (98%) with lower cell death, when comparing to discontinuous diafiltration. Overall, an integrated process allowed for a shorter process time, recovering 70% of viable hMSC (>95%), with no changes in terms of morphology, immunophenotype, proliferation capacity and multipotent differentiation potential.

Original languageEnglish
Pages (from-to)97-108
Number of pages12
JournalJournal of Biotechnology
Volume213
DOIs
Publication statusPublished - 10 Nov 2015

Fingerprint

Stem cells
Mesenchymal Stromal Cells
Processing
Cell death
Perfusion
Bioreactors
Metabolism
Washing
Proteins
Recovery
Cell Death
Cell Count

Keywords

  • Alternating tangential flow
  • Cell therapy
  • Mesenchymal stem cells
  • Perfusion
  • Process integration
  • Tangential flow filtration

Cite this

Cunha, Bárbara ; Aguiar, Tiago ; Silva, Marta M. ; Silva, Ricardo J S ; Sousa, Marcos F Q ; Pineda, Earl ; Peixoto, Cristina ; Carrondo, Manuel José Teixeira ; Serra, Margarida ; Alves, Paula M. / Exploring continuous and integrated strategies for the up- and downstream processing of human mesenchymal stem cells. In: Journal of Biotechnology. 2015 ; Vol. 213. pp. 97-108.
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Exploring continuous and integrated strategies for the up- and downstream processing of human mesenchymal stem cells. / Cunha, Bárbara; Aguiar, Tiago; Silva, Marta M.; Silva, Ricardo J S; Sousa, Marcos F Q; Pineda, Earl; Peixoto, Cristina; Carrondo, Manuel José Teixeira; Serra, Margarida; Alves, Paula M.

In: Journal of Biotechnology, Vol. 213, 10.11.2015, p. 97-108.

Research output: Contribution to journalArticle

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