Institute of Animal Genetics and Breeding, College of Animal Science and Technology, Sichuan Agricultural University, Wenjiang, 611130, Sichuan, People's Republic of China.
Cytotechnology. 2015 Oct;67(5):773-81. doi: 10.1007/s10616-014-9715-2. Epub 2014 Apr 3.
In the present study, we isolated preadipocytes from the adipose tissue of Peking duck and subsequently cultured them in vitro. Cell counting kit-8 assay was employed to establish the growth curve of duck primary preadipocytes. Meanwhile, after the cells reaching full confluency, they were induced to differentiate into mature adipocytes by the addition of a cocktail containing dexamethasone, insulin, 3-isobutyl-1-methylxanthine, and oleic acid for 8 days. Successful differentiation was demonstrated by the development of lipid droplets and the expression of key marker genes including peroxisome proliferator-activated receptor-γ (PPARγ), CCAAT/enhancer binding protein-α (CEBP/α) and adipocyte fatty acid-binding protein (FABP4). Our results showed that duck primary preadipocytes began to adhere 12 h after seeding as short spindle shapes or litter triangles, which grew quickly 3 days post attachment and maintained stable after day 7. After 8 days the preadipocytes were induced to differentiate into mature adipocytes, which were stained red by oil red O. Additionally, it showed that during preadipocyte differentiation PPARγ mRNA was highly expressed at day 3, while CEBP/α and FABP4 mRNA peaked at day 5 and 8, respectively. These results indicate that we have successfully isolated and cultured Peking duck preadipocytes and successfully induced them to differentiate into mature adipocytes. This work could lay a foundation for further research into waterfowl adipogenesis.
在本研究中，我们从北京鸭的脂肪组织中分离出前体脂肪细胞，然后在体外进行培养。使用细胞计数试剂盒-8 测定法建立了鸭原代前体脂肪细胞的生长曲线。同时，当细胞达到完全汇合后，通过添加包含地塞米松、胰岛素、3-异丁基-1-甲基黄嘌呤和油酸的鸡尾酒诱导它们分化为成熟的脂肪细胞，诱导时间为 8 天。成功分化的标志是脂滴的发育和关键标记基因的表达，包括过氧化物酶体增殖物激活受体-γ (PPARγ)、CCAAT/增强子结合蛋白-α (CEBP/α) 和脂肪细胞脂肪酸结合蛋白 (FABP4)。我们的结果表明，鸭原代前体脂肪细胞在接种后 12 小时开始贴壁，呈短梭形或小三角形，附着后 3 天快速生长，第 7 天以后保持稳定。8 天后，前体脂肪细胞被诱导分化为成熟脂肪细胞，用油红 O 染色呈红色。此外，研究结果显示，在前体脂肪细胞分化过程中，PPARγ mRNA 在第 3 天表达量较高，而 CEBP/α 和 FABP4 mRNA 在第 5 天和第 8 天分别达到峰值。这些结果表明，我们已经成功地分离和培养了北京鸭前体脂肪细胞，并成功地诱导它们分化为成熟的脂肪细胞。这项工作为进一步研究水禽脂肪生成奠定了基础。
