Multicolor staining of globin subtypes reveals impaired globin switching during erythropoiesis in human pluripotent stem cells.

ABSTRACT: Adult hemoglobin composed of α- and β-globin reflects a change from expression of embryonic ε- and fetal γ-globin to adult β-globin in human erythroid cells, so-called globin switching. Human pluripotent stem cells (hPSCs) are a potential source for in vitro erythrocyte production, but they show prominent expression of γ-globin with little β-globin expression, which indicates incomplete globin switching. To examine the mechanism of this impaired globin switching, we optimized multicolor flow cytometry to simultaneously follow expression of different globin subtypes using different immunofluorescent probes. This enabled us to detect upregulation of β-globin and the corresponding silencing of γ-globin at the single-cell level during cord blood CD34(+) cell-derived erythropoiesis, examined as

SUBMITTER: Ochi K 

PROVIDER: S-EPMC4073826 | biostudies-literature | 2014 Jul

REPOSITORIES: biostudies-literature