Early Intervention for Spinal Cord Injury with Human Induced Pluripotent Stem Cells Oligodendrocyte Progenitors
Fig 4
Indirect immunofluorescence marker characterization of oligodendrocyte progenitors derived from human iPS cells.
Nuclei were stained with DAPI (blue). Results show reduced expression of early markers (A) A2B5 and (B) NG2 and increased protein expression of mid to late oligodendrocyte progenitor (OP) differentiation markers (C) O4 (D) RIP (E) O1 (F) myelin-associated glycoprotein (MAG) and (G) myelin-associated oligodendrocytic basic protein (MOBP). Myelin basic protein (MBP) expression was not detectable by immunostaining. 1% or less of OP cells expressed markers of other neural lineages as denoted by (H) the neuronal maker neuron-specific class III beta-tubulin (TUJ1) and (I) the astrocyte marker glial fibrillary acidic protein (GFAP). (J) Quantification of the indirect immunofluorescence analyses calculating the percent of cells expressing a particular marker is shown in the lower panel. A higher percent of cells derived from BC1 expressed later OP markers MAG, MOBP and O1 while fewer cells from the BC1 lines expressed early neural progenitor (NP) and glial progenitor (GP) markers and early OP marker O4 compared to MR31 and A1-4. Differentiation of each cell line was performed in triplicate and the percent of positively stained cells was determined from 3 randomly, chosen 10X fields per 24-well cell culture dish from all three replicates. Values in the graphs represent mean ± SEM. * and † denote statistical significance compared to BC1 and MR31, respectively by ANOVA and Tukey post hoc tests (N = 9, P < 0.001). Scale bars: 100 μm.