Expansion of bone marrow and skeletal muscle SP cells in suspension.

<p>A – Representative FACS plots of P2 bone marrow suspension culture analyses for EGFP-fluorescence at days 3, 10, and 15 (left). Plots show 20,000 events. Day 15 cells were assessed for Hoechst dye effusion (right). Plots show 50,000 events. B – FACS plots of P2 skeletal muscle culture analyses for EGFP-fluorescence at days 3, 10, and 15 (left). Day 15 cells were assessed for Hoechst dye effusion (right). Both bone marrow and skeletal muscle expansion cultures demonstrated an increase in EGFP⁺ cells over time and many cells were Hoechst^low, analogous to freshly-isolated SP cells. C – Expansion of EGFP⁺ cells in P0 stirred-flask cultures as determined by hemocytometry. Numerical values for the total number of EGFP⁺ cells are expressed as Mean ± SD (<i>n</i> = 4) and asterisks represent statistical significance (* P<0.05 and ** P<0.01) compared to day 3 values. D – Expansion of bone marrow (left) and skeletal muscle (right) EGFP⁺ cells over the course of 3 passages (P0, P1, and P2). The percentage of EGFP⁺ cells in the suspension cultures is expressed as Mean ± SD (<i>n</i> = 4) and asterisks represent statistical significance (* P<0.05 and ** P<0.01) compared to day 3 values.