Improving washing strategies of human mesenchymal stem cells using negative mode expanded bed chromatography

Bárbara Cunha, Ricardo J S Silva, Tiago Aguiar, Margarida Serra, John Daicic, Jean Luc Maloisel, John Clachan, Anna Åkerblom, Manuel J T Carrondo, Cristina Peixoto, Paula M. Alves

Research output: Contribution to journalArticle

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Abstract

The use of human mesenchymal stem cells (hMSC) in clinical applications has been increasing over the last decade. However, to be applied in a clinical setting hMSC need to comply with specific requirements in terms of identity, potency and purity.This study reports the improvement of established tangential flow filtration (TFF)-based washing strategies, further increasing hMSC purity, using negative mode expanded bed adsorption (EBA) chromatography with a new multimodal prototype matrix based on core-shell bead technology.The matrix was characterized and a stable, expanded bed could be obtained using standard equipment adapted from what is used for conventional packed bed chromatography processes. The effect of different expansion rates on cell recovery yield and protein removal capacity was assessed. The best trade-off between cell recovery (89%) and protein clearance (67%) was achieved using an intermediate expansion bed rate (1.4).Furthermore, we also showed that EBA chromatography can be efficiently integrated on the already established process for the downstream processing (DSP) of hMSC, where it improved the washing efficiency more than 10-fold, recovering approximately 70% of cells after global processing. This strategy showed not to impact cell viability (> 95%), neither hMSC's characteristics in terms of morphology, immunophenotype, proliferation, adhesion capacity and multipotent differentiation potential.

Original languageEnglish
Pages (from-to)292-303
Number of pages12
JournalJournal Of Chromatography A
Volume1429
DOIs
Publication statusPublished - 15 Jan 2016

Fingerprint

Chromatography
Stem cells
Mesenchymal Stromal Cells
Washing
Adsorption
Recovery
Packed beds
Processing
Cell Survival
Proteins
Adhesion
Cells
Technology
Equipment and Supplies

Keywords

  • Cell purification
  • Cell washing
  • Downstream processing
  • Expanded bed chromatography
  • Human mesenchymal stem cells

Cite this

Cunha, Bárbara ; Silva, Ricardo J S ; Aguiar, Tiago ; Serra, Margarida ; Daicic, John ; Maloisel, Jean Luc ; Clachan, John ; Åkerblom, Anna ; Carrondo, Manuel J T ; Peixoto, Cristina ; Alves, Paula M. / Improving washing strategies of human mesenchymal stem cells using negative mode expanded bed chromatography. In: Journal Of Chromatography A. 2016 ; Vol. 1429. pp. 292-303.
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Improving washing strategies of human mesenchymal stem cells using negative mode expanded bed chromatography. / Cunha, Bárbara; Silva, Ricardo J S; Aguiar, Tiago; Serra, Margarida; Daicic, John; Maloisel, Jean Luc; Clachan, John; Åkerblom, Anna; Carrondo, Manuel J T; Peixoto, Cristina; Alves, Paula M.

In: Journal Of Chromatography A, Vol. 1429, 15.01.2016, p. 292-303.

Research output: Contribution to journalArticle

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AU - Cunha, Bárbara

AU - Silva, Ricardo J S

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AU - Maloisel, Jean Luc

AU - Clachan, John

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AU - Alves, Paula M.

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