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. 2021 Nov 4;10(11):3021. doi: 10.3390/cells10113021

Figure 1.

Figure 1

In vitro characterization of the transplants comprising aNSCs from PVRV-SVZ or from PVRMB. (A,B) aNSCs from PVRV-SVZ and PVRMB were expanded for 14 days and analyzed by immunocytochemistry. (A) Representative fluorescence images of NG2 (green, upper panel), Olig2 (magenta, middle panel) and Nestin (green, lower panel) of aNSCs derived from PVRV-SVZ (left column) and PVRMB (right column) are shown. (B) Both aNSC types retained an undifferentiated progenitor cell state with a high expression of Nestin and oligodendroglial progenitor cell markers (NG2, Olig2) with no significant differences between aNSCs derived from the PVRV-SVZ and PVRMB. (C,D) After the terminal differentiation of aNSCs for 14 days, we found small amounts of undifferentiated aNSCs and an overall astroglial commitment with a majority of GFAP+ astrocytes and smaller numbers of Tuj1+ neurons. Representative fluorescence images of Nestin (magenta, upper panel), tubulin III (Tuj1, green, middle panel) and GFAP (white, lower panel) of terminally differentiated aNSCs cultures derived from PVRV-SVZ (right column) and PVRMB (left column). (D) The quantification of undifferentiated Nestin+ cells as well as the astroglial and neuronal differentiation capacities revealed no significant differences between aNSCs derived from the PVRV-SVZ and PVRMB. (E) Quantification of aNSCs expressing the proliferation marker Ki67 during proliferation and after differentiation (representative Ki67 stainings in (C), lower panel; the data during proliferation are from [9,39] using identical culture conditions as compared to the present study). We did not detect differences in the proliferation capacity between the two aNSC types in both culture conditions. Cell nuclei are counterstained with DAPI. Scale bars, 50 µm. All p-values are from unpaired two-sided t-tests (n = 3–7). Abbreviations: PVRV-SVZ—periventricular region of the ventricular-subventricular zone of the lateral ventricles; PVRMB—periventricular region of the mdbrain; aNSCs—adult neural stem cells; NG2—neural/glial antigen 2; Olig2—oligodendrocyte transcription factor 2; Tuj1—class III β-tubulin; GFAP—glial fibrillary acidic protein; DAPI—4′,6-diamidino-2-phenylindole.