摘要目的 鉴定人眼轮匝肌来源骨骼肌干细胞多向分化潜能.方法 取7例重睑术后患者的人睑板前眼轮匝肌,经Ⅰ型胶原酶消化和差速贴壁分离眼轮匝肌来源骨骼肌干细胞,流式细胞仪测定相关表面抗原表达.体外诱导眼轮匝肌来源骨骼肌干细胞鉴定其多向分化潜能.结果 眼轮匝肌来源骨骼肌干细胞能够贴壁生长,呈现成纤维细胞样形态,表达骨骼肌来源干细胞相关表面抗原;并能体外诱导分化为脂肪细胞、成骨细胞和软骨细胞.其成肌肉分化时可融合形成肌管样结构,并表达成肌分化特异性标记Pax3、Pax7、Myf5和MyoD.结论 人眼轮匝肌组织中含有较强成肌分化潜能的骨骼肌干细胞,有望在肌肉组织再生和疾病治疗方面获得进一步的应用.

abstractsObjective To establish the isolation and culture methods of skeletal muscle stem cells,derived from human orbicularis oculi muscle (OOMSCs),and to identify their multi-directional differentiation potential in vitro.Methods The cellswere isolated from pretarsal orbicularis oculi muscle (OOM),obtained in routine blepharoplasty surgery.The tissue was digested using collagenase type I combined with re-plating methods.Specific cell surface antigen markers were detected using flow cytometry analysis.OOMSCs were cultured under different inductive conditions,to identify their pluripotent differentiation ability.Results OOMSCs exhibited similar fibroblast-like morphology as mesenchymal stem cells with high expression of skeletal muscle-derived stem cell surface markers.OOMSCs were able to differentiate into adipocytes,osteoblasts and chondrocytes in vitro,in the presence of lineage-specific inductive media.Moreover,after myogenic induction,the differentiated cells were fused into multinucleated myotube-like structure,and positive for myogenic-related marks,Pax3,Pax7,Myf5 and MyoD.Conclusions Muscle-derived stem cells can be isolated from human OOM with myogenic differentiation properties,showing further applications potential intissue regeneration and medical therapies of muscle diseases.