FIG. 1.

Combination of coculture and growth factors enhances expansion of CD34⁺ CD133⁺ cord blood cells. Purified CD34⁺ CD133⁺ cord blood cells were cocultured with MSC-A5 or MSC-GFP in the presence of SCF, FGF, IGFBP2, TPO, and heparin or cultured in the FCF culture. (A) Coculture with MSC-A5 was observed at indicated time points using an inverted microscope (10 × magnification). (B–E) Nonadherent cells in MSC-A5 and MSC-GFP cocultures and the FCF culture were harvested at 7, 11, and 14 days, counted, and analyzed for CD34 and CD133 expression. (B) Representative CD34 versus CD133 staining profiles of cultured cells. The number indicates the percentage of DP cells in the gated region. (C) The average of percentages of DP cells in the cultures at different time points from 3 experiments. (D) Fold increase of total hematopoietic cells during the course of the cultures. (E) Fold increase of CD34⁺ CD133⁺ DP cells during the course of the cultures. Representative data from 1 of 4 experiments (Supplementary Table S1) are shown. DP, double positive; FCF, feeder cell-free; FGF, fibroblast growth factor-1; IGFBP2, insulin-like growth factor binding protein 2; MSC, mesenchymal stem cell; SCF, stem cell factor; TPO, thrombopoietin.