Benedetti Anna, Cera Gianluca, De Meo Daniele, Villani Ciro, Bouche Marina, Lozanoska-Ochser Biliana
Department of Anatomical, Histological, Forensic and Orthopedic Sciences, Section of Histology and Embryology, Sapienza University of Rome, Rome, Italy.
Department of Anatomical, Histological, Forensic and Orthopedic Sciences, Section of Orthopedics, Sapienza University of Rome, Rome, Italy.
Bio Protoc. 2021 Dec 5;11(23):e4238. doi: 10.21769/BioProtoc.4238.
Satellite cells (SCs) are muscle stem cells capable of regenerating injured muscle. The study of their functional potential depends on the availability of methods for the isolation and expansion of pure SCs, which retain myogenic properties after serial passages . Here, we describe a protocol for the isolation and expansion of highly pure mouse and human SCs based on ice-cold treatment (ICT). The ICT is carried out by briefly incubating the dish containing a heterogeneous mix of adherent muscle mononuclear cells on ice for 15-30 min, which leads to the detachment only of the SCs, and gives rise to SC cultures with 95-100% purity. This approach can also be used to passage the cells, allowing SC expansion over extended periods of time without compromising their proliferation or differentiation potential. Overall, the ICT method is cost-effective, accessible, technically simple, reproducible, and highly efficient. Graphic abstract: Figure 1.Satellite cell isolation using the ice-cold treatment method.
卫星细胞（SCs）是能够再生受损肌肉的肌肉干细胞。对其功能潜能的研究依赖于分离和扩增纯卫星细胞方法的可用性，这些卫星细胞在连续传代后仍保留成肌特性。在此，我们描述了一种基于冰冷处理（ICT）分离和扩增高度纯化的小鼠和人类卫星细胞的方案。冰冷处理是通过将含有贴壁肌肉单核细胞异质混合物的培养皿在冰上短暂孵育15 - 30分钟来进行的，这只会导致卫星细胞脱离，并产生纯度为95 - 100%的卫星细胞培养物。这种方法也可用于传代细胞，使卫星细胞能够长时间扩增而不影响其增殖或分化潜能。总体而言，冰冷处理方法具有成本效益、易于操作、技术简单、可重复且高效的特点。图形摘要：图1. 使用冰冷处理方法分离卫星细胞。
