Chen Yu-Jen, Liu Hui-Yu, Chang Yun-Tsui, Cheng Ying-Hung, Mersmann Harry J, Kuo Wen-Hung, Ding Shih-Torng
Institute of Biotechnology, National Taiwan University; Department of Animal Science and Technology, National Taiwan University.
Department of Animal Science and Technology, National Taiwan University.
J Vis Exp. 2016 Mar 31(109):e53886. doi: 10.3791/53886.
Obesity is an unconstrained worldwide epidemic. Unraveling molecular controls in adipose tissue development holds promise to treat obesity or diabetes. Although numerous immortalized adipogenic cell lines have been established, adipose-derived stem cells from the stromal vascular fraction of subcutaneous white adipose tissues provide a reliable cellular system ex vivo much closer to adipose development in vivo. Pig adipose-derived stem cells (pADSC) are isolated from 7- to 9-day old piglets. The dorsal white fat depot of porcine subcutaneous adipose tissues is sliced, minced and collagenase digested. These pADSC exhibit strong potential to differentiate into adipocytes. Moreover, the pADSC also possess multipotency, assessed by selective stem cell markers, to differentiate into various mesenchymal cell types including adipocytes, osteocytes, and chondrocytes. These pADSC can be used for clarification of molecular switches in regulating classical adipocyte differentiation or in direction to other mesenchymal cell types of mesodermal origin. Furthermore, extended lineages into cells of ectodermal and endodermal origin have recently been achieved. Therefore, pADSC derived in this protocol provide an abundant and assessable source of adult mesenchymal stem cells with full multipotency for studying adipose development and application to tissue engineering of regenerative medicine.
肥胖是一种不受控制的全球性流行病。揭示脂肪组织发育中的分子调控有望治疗肥胖症或糖尿病。尽管已经建立了许多永生化的脂肪生成细胞系，但来自皮下白色脂肪组织基质血管部分的脂肪来源干细胞在体外提供了一个更接近体内脂肪发育的可靠细胞系统。猪脂肪来源干细胞（pADSC）从7至9日龄的仔猪中分离得到。将猪皮下脂肪组织的背部白色脂肪库切片、切碎并进行胶原酶消化。这些pADSC具有很强的分化为脂肪细胞的潜力。此外，通过选择性干细胞标志物评估，pADSC还具有多能性，可分化为包括脂肪细胞、骨细胞和软骨细胞在内的各种间充质细胞类型。这些pADSC可用于阐明调节经典脂肪细胞分化或向其他中胚层来源的间充质细胞类型分化的分子开关。此外，最近已经实现了向外胚层和内胚层来源细胞的扩展谱系。因此，本方案中获得的pADSC为研究脂肪发育和应用于再生医学组织工程提供了丰富且可获取的具有完全多能性的成人间充质干细胞来源。
