Murine Long-Term Repopulating Ability Is Compromised by Ex Vivo Culture in Serum-Free Medium Despite Preservation of Committed Progenitors

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Abstract
Hematopoietic progenitor cells can be maintained and expanded ex vivo in standard or serum-free culture medium supplemented with a variety of stimulatory cytokines. The use of serum-free medium allows specification of reproducible and precise growth conditions optimal for various applications and is more acceptable from a safety and regulatory point of view. Human and murine committed progenitor cells have been shown to be equivalently or better supported by serum-free culture conditions in the presence of multicytokine combinations, but there is little information on the effects of such culture conditions on repopulating stem cells. We used a murine competitive repopulation model to assess the effect of serum-free versus serum-containing ex vivo culture on long-term reconstituting cells. Despite equivalent numbers of committed CFU-C and day 12 CFU-S present after 4 days of culture of murine marrow in serum-free or serum-containing conditions in the presence of IL-3, IL-6, and SCF, long-term reconstituting activity was significantly impaired by serum-free culture. These findings may have important implications for transplantation and gene therapy applications.