Histone Deacetylase Inhibition Enhances Self Renewal and Cardioprotection by Human Cord Blood-Derived CD34+ Cells
Figure 2
Analysis of CD34+ cells proliferation in the presence and the absence of VPA by flow cytometry.
(A) CFSE staining profiles of control and VPA-treated cells at 5 and 7 days of culture. Note that the fluorescence intensity reduction as a consequence in cell proliferation was less pronounced in VPA vs. CTR cells at both time points. Proliferation index at 7 days was significantly reduced. (B) Seven days VPA treated cells had a higher frequency of slow dividing immature (CD34bright) stem cells (blue area in contour plots), as detected by co-staining with CFSE and CD34 antibody; plots on the left indicate the fluorescence profile of cells stained with CFSE and CD34 isotype antibody (iso). (C) Cell cycle analysis in 7 days CTR and VPA-treated cells revealed a higher frequency of cells in the G0–G1 and a lower percentage in either S and G2-M phases. (D) The percentage of cells specifically arrested in G0 was also increased, as detected by co-staining with anti Ki67 and CD34 antibodies at the same time point. * indicate P<0.05 by paired t-test (n≥3). (E) Quantification of the relative expression level (2−ΔΔCt method) of small cyclin/CDK inhibitors (p14ARF, p16INK4, p21Cip1/waf1 and p27) by qRT-PCR analysis. * indicates P<0.05 by unpaired t-test (n≥3).