摘要
背景:近年来间充质干细胞在骨与关节损伤治疗中的价值得到广泛认可,但目前间充质干细胞的来源较为有限,多局限在骨髓、脂肪等。目的:从大鼠的筋膜组织分离出间充质干细胞,通过研究筋膜来源的间充质干细胞生长特性及多向分化能力,以期为软骨及软组织损伤修复提供更多的细胞来源。方法:取大鼠后肢臀大肌表面的筋膜组织及膝关节软骨分别做原代细胞培养,利用流式细胞仪检测筋膜源性干细胞表面分子表达,利用成脂诱导分化培养基、成软骨诱导分化培养基诱导其多向分化,并进行相应检测。结果:流式细胞仪检测显示筋膜来源的干细胞CD29、CD90、CD71阳性表达,CD45阴性表达;成脂诱导分化油红O染色呈阳性,3D培养的细胞团块透亮,与正常软骨形态相近,阿尔新蓝、番红O染色以及免疫组化显示3D诱导分化培养的细胞团块特异性合成Ⅱ型胶原和糖胺聚糖。结论:筋膜来源的间充质干细胞具有较强的成软骨分化能力,可以为间充质干细胞治疗骨与关节损伤提供可选择的细胞来源。
Background:In recent years mesenchymal stem cell is widely known as one treatment for bone and joint injury,but the source of mesenchymal stem cell is limited at spine marrow and fat.Objective:To isolate the mesenchymal stem cells from the fascia of the left gluteus maximus in rat and to identify the growth features and multi-directional differentiation ability of fascial derived mesenchymal stem cells in order to provide more source for cartilage regeneration and repair.Methods:Fascia isolated from the left gluteus maximus and cartilages isolated from knee joint of SD rats were taken for primary cell culture.Cell surface markers were checked by flow cytometry;it was induced for multi-directional differentiation through chondrogenesis and adipogenesis cultures;related detections were conducted.Results:Fascial-derived stem cells expressed CD29,CD71 and CD90 as detected by flow cytometry,but CD45 was not detected by flow cytometry.Oil red O staining was positive in the adipogenic-induction.3 D cultured cell aggregate was lucency transparent which was similar with the normal cartilage and could specifically synthesize collagenⅡand glycosaminoglycan showed by Alcian blue and Sarranine O stainings and immunohistochemistry test.Conclusions:Rat fascial-derived mesenchymal stem cell has strong differentiation ability into chondrocytes and fascia may become a new source of mesenchymal stem cell in treating bone and joint injury.
作者
史简铭
田进翔
辛超飞
龚汉普
常英健
李广恒
许建中
SHI Jianming;TIAN Jinxiang;XIN Chaofei;GONG Hanpu;CHANG Yingjian;LI Guangheng;XU Jianzhong(Department of Orthopedics,The First Affiliated Hospital of Zhengzhou University,Zhengzhou 450052,China)
出处
《中华骨与关节外科杂志》
2019年第8期625-630,共6页
Chinese Journal of Bone and Joint Surgery
关键词
筋膜间充质干细胞
成软骨诱导分化
3D培养
流式细胞检测
Fascial Derived Mesenchymal Stem Cell
Chondrogenic Induced Differentiation
3D Culture
Flow Cytometry
