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Controlling the Effective Oxygen Tension Experienced by Cells Using a Dynamic Culture Technique for Hematopoietic Ex Vivo Expansion

journal contribution
posted on 2023-02-09, 03:10 authored by A Tiwari, C S Wong, L P Nekkanti, J A Deane, C McDonald, J Li, Y Pham, A E Sutherland, G Jenkin, M A Kirkland
Clinical hematopoietic stem/progenitor cell (HSPC) transplantation outcomes are strongly correlated with the number of cells infused. Hence, to generate sufficient HSPCs for transplantation, the best culture parameters for expansion are critical. It is generally assumed that the defined oxygen (O2) set for the incubator reflects the pericellular O2 to which cells are being exposed. Studies have shown that low O2 tension maintains an undifferentiated state, but the expansion rate may be constrained because of limited diffusion in a static culture system. A combination of low ambient O2 and dynamic culture conditions has been developed to increase the reconstituting capacity of human HSPCs. In this unit, the protocols for serum-free expansion of HSPCs at 5% and 20% O2 in static and dynamic nutrient flow mode are described. Finally, the impact of O2 tension on HSPC expansion in vitro by flow cytometry and colony forming assays and in vivo through engraftment using a murine model is assessed. © 2018 by John Wiley & Sons, Inc.

History

Journal

Current Protocols in Stem Cell Biology

Volume

44

Pagination

2A.11.1 - 2A.11.13

ISSN

1941-7322

eISSN

1938-8969

Publication classification

C1 Refereed article in a scholarly journal

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