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 language | English |
|---|---|
| Pages (from-to) | 97-108 |
| Number of pages | 12 |
| Journal | Journal of Biotechnology |
| Volume | 213 |
| DOIs | |
| Publication status | Published - 10 Nov 2015 |
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Keywords
- Alternating tangential flow
- Cell therapy
- Mesenchymal stem cells
- Perfusion
- Process integration
- Tangential flow filtration
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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 journal › Article
TY - JOUR
T1 - Exploring continuous and integrated strategies for the up- and downstream processing of human mesenchymal stem cells
AU - Cunha, Bárbara
AU - Aguiar, Tiago
AU - Silva, Marta M.
AU - Silva, Ricardo J S
AU - Sousa, Marcos F Q
AU - Pineda, Earl
AU - Peixoto, Cristina
AU - Carrondo, Manuel José Teixeira
AU - Serra, Margarida
AU - Alves, Paula M.
PY - 2015/11/10
Y1 - 2015/11/10
N2 - 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.
AB - 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.
KW - Alternating tangential flow
KW - Cell therapy
KW - Mesenchymal stem cells
KW - Perfusion
KW - Process integration
KW - Tangential flow filtration
UR - http://www.scopus.com/inward/record.url?scp=84942829160&partnerID=8YFLogxK
U2 - 10.1016/j.jbiotec.2015.02.023
DO - 10.1016/j.jbiotec.2015.02.023
M3 - Article
VL - 213
SP - 97
EP - 108
JO - Journal of Biotechnology
JF - Journal of Biotechnology
SN - 0168-1656
ER -