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Isolation, culture and differentiation of duck (Anas platyrhynchos) preadipocytes

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Abstract

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.

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Abbreviations

CCK-8:

Cell counting kit-8

CEBP/α:

CCAAT/enhancer binding protein-α

DEX:

Dexamethasone

DNL:

De novo lipogenesis

FABP4:

Adipocyte fatty acid-binding protein

FBS:

Fetal bovine serum

IBMX:

3-Isobutyl-1-methylxanthine

LPL:

Lipoprotein lipase

MSCs:

Mesenchymal stem cells

PPARγ:

Peroxisome proliferator-activated receptor-γ

SD:

Standard deviation

TG:

Triacylglycerol

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Acknowledgments

This work was supported by the earmarked fund for China Agriculture Research Service (No. CARS-43-6).

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Authors and Affiliations

  1. Institute of Animal Genetics and Breeding, College of Animal Science and Technology, Sichuan Agricultural University, Wenjiang, 611130, Sichuan, People’s Republic of China

    Fang Ding, Qing-qing Li, Le Li, Chao Gan, Xin Yuan, Hua Gou, Hua He, Chun-chun Han & Ji-wen Wang

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  1. Fang Ding
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  2. Qing-qing Li
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  3. Le Li
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  4. Chao Gan
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  5. Xin Yuan
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  6. Hua Gou
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  7. Hua He
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  8. Chun-chun Han
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  9. Ji-wen Wang
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Correspondence to Ji-wen Wang.

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Ding, F., Li, Qq., Li, L. et al. Isolation, culture and differentiation of duck (Anas platyrhynchos) preadipocytes. Cytotechnology 67, 773–781 (2015). https://doi.org/10.1007/s10616-014-9715-2

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  • DOI: https://doi.org/10.1007/s10616-014-9715-2

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