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

Joan Sebastián Gallego-Murillo, Giulia Iacono, Luuk A.M. van der Wielen, Emile van den Akker, Marieke von Lindern*, Sebastian Aljoscha Wahl

*Corresponding author for this work

Research output: Contribution to journalArticleScientificpeer-review

5 Citations (Scopus)
83 Downloads (Pure)

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.

Original languageEnglish
Pages (from-to)3096-3116
JournalBiotechnology and Bioengineering
Volume119
Issue number11
DOIs
Publication statusPublished - 2022

Keywords

  • cell culture
  • cultured blood
  • erythropoiesis
  • red blood cell
  • scale-up
  • stirred tank bioreactor

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