Small Molecule-Assisted, Line-Independent Maintenance of Human Pluripotent Stem Cells in Defined Conditions
Figure 1
Dorsomorphin prevents spontaneous TE-like differentiation in chemically defined hESC medium.
(A) Colonies in N2B27+ FGF2 fail to expand well after several passages (line HuES6). (B) Typical spontaneous differentiation formed in centers of colonies. Representative differentiated and undifferentiated parts were collected for further analysis (right panels). (C) Real-time RT-PCR analysis of samples in (B). (D) Immunocytochemistry confirms that inner differentiated cells stain positive for HAND1. (E) Dorsomorphin addition to N2B27 completely prevents TE-like differentiation in routine culture (line H7). (F) Titration of optimal dorsomorphin dosage. At 50 nM, both TE-like and neural differentiation rates were minimal.