Fig. 2. Development of adipocytes in primary cultures of quail cephalic NC cells (CNCC). (A,B) Primary cultures of CNCC were obtained from mes-rhombencephalon of HH stage 9 quail embryo in explant culture and expanded for 5 days in cloning medium. Adipocyte differentiation was then induced using different media (cloning, L1 and hmads) and adipogenic treatments (DIF1 or DIF2). Adipocytes were identified after 15 days. (A) Typical adipocytes show lipid droplet-filled cytoplasm (left) and are stained with Oil Red O, which reveals neutral lipids (right). Scale bar: 100 µm. (B) Quantification of CNCC primary cultures containing adipocytes after treatment with the mentioned media and adipogenic treatments. A total of 100 cultures were analysed. (C,D) Secondary cultures of quail CNCC were isolated from 48-hour primary cultures. Adipocyte differentiation was then induced at day 6 using the mentioned media and adipogenic treatments. (C) Quantification of secondary CNCC cultures containing adipocytes after 15 days of treatment. A total of 84 cultures were analysed. (D) Expression of CEBP, PPAR, FABP4 and 18s RNAs after 3 days in cloning medium+DIF1. The results are shown for two independent CNCC cultures out of 10 distinct experiments.