TY - JOUR
T1 - Improving washing strategies of human mesenchymal stem cells using negative mode expanded bed chromatography
AU - Cunha, Bárbara
AU - Silva, Ricardo J S
AU - Aguiar, Tiago
AU - Serra, Margarida
AU - Daicic, John
AU - Maloisel, Jean Luc
AU - Clachan, John
AU - Åkerblom, Anna
AU - Carrondo, Manuel J T
AU - Peixoto, Cristina
AU - Alves, Paula M.
PY - 2016/1/15
Y1 - 2016/1/15
N2 - 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.
AB - 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.
KW - Cell purification
KW - Cell washing
KW - Downstream processing
KW - Expanded bed chromatography
KW - Human mesenchymal stem cells
UR - http://www.scopus.com/inward/record.url?scp=84954198315&partnerID=8YFLogxK
U2 - 10.1016/j.chroma.2015.12.052
DO - 10.1016/j.chroma.2015.12.052
M3 - Article
C2 - 26739915
AN - SCOPUS:84954198315
SN - 0021-9673
VL - 1429
SP - 292
EP - 303
JO - Journal Of Chromatography A
JF - Journal Of Chromatography A
ER -