Isolation and identification of chicken adipose-derived mesenchymal stem cells

【Objective】 A method for isolation and identification of chicken adipose-derived mesenchymal stem cells(AMSCs) was established.【Method】 AMSCs from Tianlu yellow chickens were obtained by type I collagenase digestion. CCK-8 was used to detect cell activities. RT-PCR was used to examine their specific marker. Whereas their adipogenic and osteogenic differentiations were chemically induced.【Result and conclusion】The primary cultured and subcultured cells showed fibroblast-like morphology, and primary AMSCs were subcultured to passage 10 without any change in activities. The growth curves were typically sigmoidal. RT-PCR assays showed that the specific markers of AMSCs, CD29, CD44 and CD71, were positive, but CD34 and CD45 characterized by hematopoietic stem cells were negative. In addition, AMSCs can successfully differentiated into osteoblasts and adipocytes in different media. Lipid droplets formation was recorded during adipogenic induction, with the cells being positive for oil red O staining, and mRNA expression of peroxisome proliferator-activated receptor-γ(PPARγ) and fatty acid（FAS）was increased. During osteogenic induction, alizarin red staining showed that the calcium nodus was positive, and there was significant difference in alkaline phosphatase (ALP) activities between the test and control group (P<0.05), mRNA expression of ALP and bone morphogenetic protein-2 (BMP2) also increased. This research suggests that the mesenchymal stem cells isolated from chicken adipose tissues are multi potential and may provide the possibility for future clinical application.