Expansion and differentiation of ex vivo cultured erythroblasts in scalable stirred bioreactors

Journal Article (2022)
Author(s)

Joan S. Gallego Murillo (TU Delft - BT/Bioprocess Engineering)

Giulia Iacono (Amsterdam UMC)

Van Der Wielen Luuk (University of Limerick, TU Delft - BT/Bioprocess Engineering)

Emile van den Akker (Amsterdam UMC)

Marieke von Lindern (Amsterdam UMC)

Aljoscha Wahl (TU Delft - BT/Industriele Microbiologie)

Research Group
BT/Bioprocess Engineering
Copyright
© 2022 Joan Sebastián Gallego Murillo, Giulia Iacono, L.A.M. van der Wielen, Emile van den Akker, Marieke von Lindern, S.A. Wahl
DOI related publication
https://doi.org/10.1002/bit.28193
More Info
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Publication Year
2022
Language
English
Copyright
© 2022 Joan Sebastián Gallego Murillo, Giulia Iacono, L.A.M. van der Wielen, Emile van den Akker, Marieke von Lindern, S.A. Wahl
Related content
Research Group
BT/Bioprocess Engineering
Issue number
11
Volume number
119
Pages (from-to)
3096-3116
Reuse Rights

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Abstract

Transfusion of donor-derived red blood cells (RBCs) is the most common form of cell therapy. Production of transfusion-ready cultured RBCs (cRBCs) is a promising replacement for the current, fully donor-dependent therapy. A single transfusion unit, however, contains 2 × 1012 RBC, which requires large scale production. Here, we report on the scale-up of cRBC production from static cultures of erythroblasts to 3 L stirred tank bioreactors, and identify the effect of operating conditions on the efficiency of the process. Oxygen requirement of proliferating erythroblasts (0.55–2.01 pg/cell/h) required sparging of air to maintain the dissolved oxygen concentration at the tested setpoint (2.88 mg O2/L). Erythroblasts could be cultured at dissolved oxygen concentrations as low as 0.7 O2 mg/ml without negative impact on proliferation, viability or differentiation dynamics. Stirring speeds of up to 600 rpm supported erythroblast proliferation, while 1800 rpm led to a transient halt in growth and accelerated differentiation followed by a recovery after 5 days of culture. Erythroblasts differentiated in bioreactors, with final enucleation levels and hemoglobin content similar to parallel cultures under static conditions.