Fig. 3.

MCDM medium maintains the first branchial arch post-migratory CNC cells in an undifferentiated state. A: Sorted CNC cells 4 hr after plating on FN-coated dish in medium II (MCDM medium). These cells show attachment to the plate. B: The undifferentiated cells, which are featured by their small body size, intense nuclei, and deep color, have reached confluence on day 4. Following immunofluorescent staining with antibodies against NF145 (C), S100 (D), or α-SMA (E), it is clear that these are undifferentiated CNC cells following 4 days of culture in medium II. Alkaline phosphatase staining with BCIP/NBT reagent is also negative (F). When MCDM is switched to DMCDM medium on day 5, morphological changes begin to take place, suggesting differentiation of CNC cells following 2 (G) or 4 (H) additional days of culture.