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

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.

The differentiation of preadipocytes and gene expression related to adipogenesis in ducks (Anas platyrhynchos)

“…Pre-adipocytes from Cherry Valley duck (lean-meat duck), White-crested duck (egg-laying duck), and Jianchang duck (fatty-meat duck) were isolated as described below [ 3 ]. Cherry Valley duck and Jianchang duck were provided by Jiangsu Yike Food Co., Ltd, and White-crested duck was provided by Waterfowl Institute of Zhenjiang City.…”

“… Note: PPARγ : peroxisome proliferator activated receptor γ, LPL : lipoprotion lipase, DLK1 : delta-like 1 homologue, C/EBPα : CCAAT/ enhancer binding protein- α , FABP4 : adipocyte fatty acid binding protein 4, FAS :fatty acid synthetase. The primer of CEBP/α , FABP4 were picked from Ding et al [ 3 ] …”

“…And the adipocytes aggregated into clusters are divided into small lobes by thin layers of loose connective tissue. All known adipocytes derive from pluripotent mesenchymal stem cells (MSC) in a process called adipogenesis [ 1 – 3 ]. The proliferation and differentiation of pre-adipocytes leads to the formation of adipose tissue.…”

Protocols and pitfalls in obtaining fatty acid-binding proteins for biophysical studies of ligand-protein and protein-protein interactions

“…Nonetheless, oligomerization state is a potentially important consideration with respect to AFABP activation, making it essential to recognize and control the formation of aggregates, including both non-covalently associated and disulfide-linked dimers. Dimers of the type observed herein could also be formed by other FABPs for which the Cys-1 residue is conserved; examples include bacterial, fruit fly, poultry, and fish homologs but no other mammalian FABPs [37] , [38] , [39] , [40] . S-S linkages in this protein family have been reported in two instances: for rat cutaneous FABP (FABP5), which has six cysteine residues and forms an intramolecular disulfide linkage (but no dimers), putatively by a red-ox mechanism [41] ; and for human heart FABP (FABP3), where dimerization secondary to an intermolecular S-S bond was found [11] .…”

Genome-wide analysis of lncRNAs and mRNAs expression during the differentiation of abdominal preadipocytes in chicken

“…The growth of adipose tissue is primarily due to the increase in adipocyte cell number (hyperplasia) and the enlargement of adipocytes (hypertrophy). As is well known that adipocytes are derived from pluripotent mesenchymal stem cells (MSCs) in adipogenesis process(Ding et al, 2015). The MSC have ability to develop into adipoblasts, which then will develop into preadipocytes that could store lipid.…”