Fig. 2
Identification of multipotent neural crest-derived progenitor cells in the carotid body. (A) Dot plot showing the expression of progenitor cell surface markers at the population level. (B) Flow cytometry plot illustrating the expression of two cell surface markers (CADM2 and HNK-1) in carotid body cells, including the gating strategy used for cell sorting. (C) Neurosphere (NS) formation by progenitor cells. Left: Percentage of sorted cells, based on each surface marker, from the total carotid body cell preparation. Center: Light microscopy images of NS formed after 10 days in culture using each cell surface marker. Arrowheads indicate typical surface differentiating blebs characteristic of carotid body NS14. Scale bar: 100 µm. Right: Efficiency of NS formation, calculated as the percentage of sorted cells that form NS. (D) Immunocytochemistry on NS cultured in adherent substrate, using antibodies against tyrosine hydroxylase (TH) and smooth muscle actin (SMA) to classify NS types. Mesenchymal NS contained only SMA + cells, neuronal NS contained only TH + cells, and multipotent NS contained both. Cells were counterstained with DAPI. Scale bars: 50 µm. Insets: 25 µm. (E) Quantification of NS subtypes obtained by sorting with each cell surface marker.
