Figure 1. Effect of chromatin modifying agents on ex vivo expansion of mobilized peripheral blood (MPB) hematopoietic stem cells.

A) Effects of 5azaD/TSA treatment on CD34 and CD90 expression following culture. Results are representative of 1 of 5 independent experiments in cytokine A condition.

B) Effect of 5azaD/TSA treatment on proportions of CD34⁺ and CD34⁺ CD90⁺ cells at Day 9 of culture in cytokine-only versus sequential 5azaD/TSA + cytokine environment. Cytokine A and B represent previously determined optimal and worst cytokine conditions for expansion of CD34⁺ CD90⁺ cord blood cells, respectively. A higher proportion of CD34⁺ CD90⁺ cells was observed in MPB cells cultured with 5azaD/TSA with either Cytokine A or B conditions compared to cells cultured in Cytokine A or B alone (Cytokine A: 15.0 ± 2.8% vs. 1.2 ± 0.4%, p =0.048, n = 5; Cytokine B: 5.7 ± 0.9% vs. 0.3 ± 0.03%, n = 2). Data represent mean ± standard errors of mean.

C) Effect of 5azaD/TSA on fold expansion of CD34⁺ CD90⁺ cells following nine days of culture. The fold expansion of CD34⁺ CD90⁺ cell numbers was determined by dividing the total numbers of viable cells expressing the phenotype at Day 9 by the input number of viable cells expressing the same phenotype at Day 0. A higher degree of expansion of CD34⁺ CD90⁺ cells was observed in MPB cells cultured with 5azaD/TSA with either Cytokine A or B conditions compared to cells cultured in Cytokine A or B alone (Cytokine A: 3.6 ± 0.5 vs. 0.7 ± 0.1; p = 0.014, n = 5; Cytokine B: 4.5 ± 0.9 vs. 0.9 ± 0.1; n = 2). Data represent mean ± standard errors of mean.