Ex-vivo expansion of hematopoietic stem/progenitor cells (HSC) would be an enormous boost towards the widespread use of umbilical cord blood (UCB) for cell therapy applications, due to the low HSC number available per UCB unit. In this study, UCB CD34+-enriched cells were co-cultured with mesenchymal stem cell (MSC)-derived feeder layers using a cytokine-supplemented serum-free medium. Using a stem cell engineering approach, important parameters affecting ex-vivo expansion performance of UCB HSC were systematically studied and optimized - serum-free media formulations and cytokine cocktails, initial CD34+ cell enrichments - resulting in an increase of cell productivity (20-fold increase in CD34+ cells after 7 days), while reducing culture costs by 50-65% compared to previously established protocols. In addition, the effect of oxygen tension in HSC-MSC co-cultures was investigated: physiological O2 levels (5-10%) were found to be beneficial for an efficient expansion of UCB HSC while cell metabolic patterns were consistent with hypoxic adaptation, being shifted to aerobic glycolysis.

Towards optimal culture conditions for the ex-vivo expansion of umbilical cord blood hematopoietic stem/progenitor cells for cell therapy

243rd National Meeting (2012)

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