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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.
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