Journal: Stem Cells International
Article Title: Isolation and Characterization of Neural Stem Cells from the Rat Inferior Colliculus
Figure Lengend Snippet: Differentiated cells from IC. NSCs after 6 days on glass coverslips in differentiation medium (DIF). (a) β -III-Tubulin (red) marked neuronal differentiated cells. The β -III-tubulin-positive cells had neuron-typical spindled, slender somata with bipolar axon configuration. (b) Astrocytes were identified by the staining of glial fibrillary acidic protein (GFAP, red). These cells showed stellate-configured branches, which is typical for astrocytes. (c) Oligodendrocytes showed positive labelling of myelin basic protein (MBP, red). Typically for oligodendrocytes, they had a peripheral-starting myelination, which is characterized by MBP staining. The proximal portions are β -tubulin-positive (green). The extensive, diversified growth of the branches is a morphological sign for oligodendrocytes. (d) Cells that were still in the undifferentiated progenitor cell stage were stained by Nestin (red). These cells had a very similar morphology to that of the GFAP-positive, astrocyte-differentiated cells. They have multipolar branches, which are labelled positively for Nestin. The cytoskeleton of all viable cells was stained by β -tubulin (green). Cell nuclei were stained with DAPI (blue).
Article Snippet: For immunocytochemistry, preparations were incubated with the following primary antibodies at 5°C for 12 h in 1% BSA solution and 0.1 M PBS buffer: mouse monoclonal against Atoh1 (1 : 1000; Ab27667, Abcam®), mouse monoclonal against BrdU (5-bromo-2′-deoxyuridine) (1 : 600; #05-633, Millipore®), mouse monoclonal against β -tubulin (1 : 1000; #TS293, Sigma-Aldrich®), mouse monoclonal against β -III-tubulin (1 : 1000; #Ab7751, Abcam®), rabbit polyclonal against β -III-tubulin (1 : 2000; #Ab18207, Abcam®), rabbit polyclonal against doublecortin (DCX) (1 : 1000; #Ab18723, Abcam®), mouse monoclonal against glial fibrillary acidic protein (GFAP) (1 : 1000; #MAB360, Millipore®), rabbit polyclonal against myelin basic protein (MBP) (1 : 800; #M3821, Sigma-Aldrich®), mouse monoclonal against Nestin (1 : 800; #MAB353, Millipore®), and rabbit polyclonal against Sox-2 (1 : 2000; #Ab97959, Abcam®).