Figure 2: MiR-219 accelerates mESC differentiation into an oligodendrocyte lineage. | Scientific Reports

Figure 2: MiR-219 accelerates mESC differentiation into an oligodendrocyte lineage.

From: Transplanted miR-219-overexpressing oligodendrocyte precursor cells promoted remyelination and improved functional recovery in a chronic demyelinated model

Figure 2

(A) A schematic protocol for the directed differentiation of miR219-mESCs into an oligodendrocyte linage. To generate OPCs, on day 15, NPCs were derived from mESCs grown for up to 28 days in DMEM/F12 supplemented with 10 ng/mL bFGF, 10 ng/mL PDGF-AA, 1% N2, and 2% KSR. Fluorescence immunocytochemistry imaging revealed that most of the GFP-positive cells in the scramble group were also A2B5 positive (B–D); a few were NG2 (E–G) and O4 (H–J) positive at 23 days in vitro. In the miR-219 group, GFP-positive cells expressed weak A2B5 (K–M) and strong NG2 (N–P) and O4 (Q–S) fluorescence intensity. Nuclei were stained with DAPI (blue). For oligodendrocyte maturation, miR219-OPCs were cultured in DMEM/F12 with 10% FBS for 7 days. Images show that OPCs differentiated into MBP- (T–U) and CNP- (V–W) positive mature oligodendrocytes. The percentages of A2B5-, NG2- and O4-positive cells in GFP-positive cells were counted (X). The data are presented as the mean ± s.e.m. *P < 0.05, **P < 0.001; n = 3 for each group. Scale bars: S = 50 μm; W = 20 μm.

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