Modification of ESC gene and surface marker expression during in vitro differentiation in fully defined medium.
(A) Oct-4 and Brachyury mRNA levels were assessed by quantitative RT-PCR at the onset of the differentiation (d0) and 4 (d4) and 8 (d8) days later. Data were normalized with Gus-B housekeeping gene expression (Representative data of 4 independent experiments). (B) CD34 expression was assessed by flow cytometry upon 8 days of differentiation before (black line, left panel) and after (black line, right panel) MACS selection. Grey profiles represent CD34 expression before hematopoietic differentiation. (C) Analysis of relevant markers was performed on MACS-enriched ES34+ after 8 days of differentiation and compared to CB34+ sorted cells. (D) SCL/Tal1 and Runx1 mRNA levels on indicated populations were assessed by quantitative RT-PCR and RT-PCR, respectively. Data are representative of 4 independent experiments.