A whole serum-deprived (WSD) medium was developed and optimized for expansion of colony-forming cells (CFC) in cord blood (CB) mononuclear cell (MNC) cultures. This medium was compared with four commercially available WSD media (commercial media), three WSD media whose compositions have been publicly disclosed (public media), two serum-containing media, and two basal media, for cell and CFC expansion in 10-day CB and mobilized peripheral blood (PB) MNC cultures supplemented with interleukin-3 (IL-3), IL-6, and stem cell factor (SCF). Selected WSD media and both serum-containing media gave significant CFC expansion in CBMNC and PBMNC cultures. The serum-containing human long-term medium gave the greatest cell (3.0-fold) and CFC (25-fold) expansions in CBMNC cultures, whereas our medium maintained the most cells (93% of input) and gave the greatest CFC expansion (6.1-fold) for PBMNC cultures. Of the commercial media, Progenitor-34 gave the greatest cell expansion (1.2-fold) and X VIVO-10 gave the greatest CFC expansion (11-fold) for CBMNC cultures, and Progenitor-34 maintained the most cells (83% of input) and gave the greatest CFC expansion (3.1-fold) for PBMNC cultures. Of the public media (including ours), our medium gave the greatest cell (1.4-fold) and CFC (6.1-fold) expansion for CBMNC cultures. Although there were slight correlations between cell and CFC expansion in 10-day CBMNC and PBMNC cultures (r2 of 0.848 and 0.594, respectively), the correlations did not give reliable predictions for medium selection. In addition, the different media favored expansion of different CFC types and performed differently for cultures using different cell sources (CB versus PB). Taken together, these results suggest that media must be carefully screened for the cell source to be cultured and the cell type(s) to be produced (e.g. total cells, CFC).
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