TY - JOUR
T1 - Bioprocess integration for human mesenchymal stem cells
T2 - From up to downstream processing scale-up to cell proteome characterization
AU - Cunha, Bárbara
AU - Aguiar, Tiago
AU - Carvalho, Sofia B.
AU - Silva, Marta M.
AU - Gomes, Ricardo A.
AU - Carrondo, Manuel J.T.
AU - Gomes-Alves, Patrícia
AU - Peixoto, Cristina
AU - Serra, Margarida
AU - Alves, Paula M.
PY - 2017/4/20
Y1 - 2017/4/20
N2 - To deliver the required cell numbers and doses to therapy, scaling-up production and purification processes (at least to the liter-scale) while maintaining cells’ characteristics is compulsory. Therefore, the aim of this work was to prove scalability of an integrated streamlined bioprocess compatible with current good manufacturing practices (cGMP) comprised by cell expansion, harvesting and volume reduction unit operations using human mesenchymal stem cells (hMSC) isolated from bone marrow (BM-MSC) and adipose tissue (AT-MSC). BM-MSC and AT-MSC expansion and harvesting steps were scaled-up from spinner flasks to 2 L scale stirred tank single-use bioreactor using synthetic microcarriers and xeno-free medium, ensuring high cellular volumetric productivities (50 × 106 cell L−1 day−1), expansion factors (14–16 fold) and cell recovery yields (80%). For the concentration step, flat sheet cassettes (FSC) and hollow fiber cartridges (HF) were compared showing a fairly linear scale-up, with a need to slightly decrease the permeate flux (30–50 LMH, respectively) to maximize cell recovery yield. Nonetheless, FSC allowed to recover 18% more cells after a volume reduction factor of 50. Overall, at the end of the entire bioprocess more than 65% of viable (>95%) hMSC could be recovered without compromising cell's critical quality attributes (CQA) of viability, identity and differentiation potential. Alongside the standard quality assays, a proteomics workflow based on mass spectrometry tools was established to characterize the impact of processing on hMSC's CQA; These analytical tools constitute a powerful tool to be used in process design and development.
AB - To deliver the required cell numbers and doses to therapy, scaling-up production and purification processes (at least to the liter-scale) while maintaining cells’ characteristics is compulsory. Therefore, the aim of this work was to prove scalability of an integrated streamlined bioprocess compatible with current good manufacturing practices (cGMP) comprised by cell expansion, harvesting and volume reduction unit operations using human mesenchymal stem cells (hMSC) isolated from bone marrow (BM-MSC) and adipose tissue (AT-MSC). BM-MSC and AT-MSC expansion and harvesting steps were scaled-up from spinner flasks to 2 L scale stirred tank single-use bioreactor using synthetic microcarriers and xeno-free medium, ensuring high cellular volumetric productivities (50 × 106 cell L−1 day−1), expansion factors (14–16 fold) and cell recovery yields (80%). For the concentration step, flat sheet cassettes (FSC) and hollow fiber cartridges (HF) were compared showing a fairly linear scale-up, with a need to slightly decrease the permeate flux (30–50 LMH, respectively) to maximize cell recovery yield. Nonetheless, FSC allowed to recover 18% more cells after a volume reduction factor of 50. Overall, at the end of the entire bioprocess more than 65% of viable (>95%) hMSC could be recovered without compromising cell's critical quality attributes (CQA) of viability, identity and differentiation potential. Alongside the standard quality assays, a proteomics workflow based on mass spectrometry tools was established to characterize the impact of processing on hMSC's CQA; These analytical tools constitute a powerful tool to be used in process design and development.
KW - Cell therapy
KW - Human mesenchymal stem cells
KW - Mass spectrometry
KW - Process development
KW - Product characterization
KW - Scale-up
UR - http://www.scopus.com/inward/record.url?scp=85014492946&partnerID=8YFLogxK
U2 - 10.1016/j.jbiotec.2017.01.014
DO - 10.1016/j.jbiotec.2017.01.014
M3 - Article
C2 - 28174039
AN - SCOPUS:85014492946
SN - 0168-1656
VL - 248
SP - 87
EP - 98
JO - Journal of Biotechnology
JF - Journal of Biotechnology
ER -