Fig. 2. Pluripotency of three types of Dnmt knockout (KO) ESCs and their differentiation potential into neural rosette and neural stem cells (NSCs). (A) Immunocytochemical analysis of control (Olig2-GFP) and three types of Dnmt KO ESCs, including Dnmt1 KO, Dnmt3a/3b double KO (Dnmt3 DKO), and Dnmt1/3a/3b triple KO (Dnmt TKO). All ESC lines expressed the pluripotency markers Oct4 and Nanog. Nuclei were counter-stained with Hoechst (blue); scale bars: 200 μm. (B) Embryoid body formation of control (Olig2-GFP) and three types of Dnmt KO ESCs; scale bars: 200 μm. (C) Neural rosette formation in control and three types of Dnmt KO ESCs. White arrows indicate neural rosettes, which stained positive for the NSC markers Sox2 and Nestin. Nuclei were counter-stained with Hoechst (blue). Control (Olig2-GFP) ESCs expressed Olig2-GFP, which is expressed in NSCs after commitment from pluripotent stem cells; scale bars: 100 μm. (D) Established NSC lines from control and three types of Dnmt KO ESCs; scale bars: 100 μm.

International Journal of Stem Cells 2023;16:44-51 https://doi.org/10.15283/ijsc22138

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