Figure 6

Hematopoietic differentiation from BiPSC13 with AID expression induced using the doxycycline-controlled (Tet-off) system. (a) Flow cytometric analysis of the cell phenotype after differentiation of BiPSC13#1-AID into hematopoietic progenitors. The population of CD34-positive cells is surrounded by a dotted line. (A) BiPSC13#1-AID were cultured in the presence of doxycycline, (B) Originally, BiPSC13#1-AID were cultured in the presence of doxycycline to inhibit expression of AID, and subsequently, doxycycline was withdrawn 10 days before sorting of the CD34-positive cells, and (C) BiPSC13#1-AID were cultured in the absence of doxycycline to express AID constitutively. (b) Immunofluorescence analysis of the expression of AID in the sorted CD34-positive cells. AID expressed cells were detected in (B) and (C) partially. Arrow heads and arrows indicate AID-positive or –negative cells, respectively. Culture condition of (A), (B), and (C) are as described in (a). (c) Wright staining of cytospins picked up from a colony (x400). Culture condition of (A), (B), and (C) are as described in (a).