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

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

•Expanded bed chromatography (EBA) was used to improve washing efficiency of hMSC.•A new multimodal matrix based on core bead technology was used in negative mode.•Expansion bed rate has impact on cell recovery yield and protein removal capacity.•Washing efficiency was improved more than 10-fold, re...

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Bibliographic Details
Published in:Journal of Chromatography A Vol. 1429; pp. 292 - 303
Main Authors: 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.
Format: Journal Article
Language:English
Published: Netherlands Elsevier B.V 15-01-2016
Subjects:
Cell purification
Cell Survival
Cell washing
Chromatography
Downstream processing
Expanded bed chromatography
Filtration
Human
Human mesenchymal stem cells
Humans
Mesenchymal Stem Cell Transplantation - methods
Mesenchymal Stromal Cells - cytology
Proteins
Purity
Recovery
Stem cells
Strategy
Washing
Expanded bed chromatography
Human mesenchymal stem cells
Cell purification
Downstream processing
Cell washing
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Summary:•Expanded bed chromatography (EBA) was used to improve washing efficiency of hMSC.•A new multimodal matrix based on core bead technology was used in negative mode.•Expansion bed rate has impact on cell recovery yield and protein removal capacity.•Washing efficiency was improved more than 10-fold, recovering 70% of cells.•After DSP, hMSC maintained their identity, potency and high cell viability. 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.
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content type line 23
ISSN:0021-9673
1873-3778
DOI:10.1016/j.chroma.2015.12.052