蒙古马脂肪来源间充质干细胞体外成脂和成骨诱导分化

Differentiation of Mongolia Horse Adipose Tissue-Derived Mesenchymal Stem Cells into Adipocytes and Osteoblasts in Vitro

取蒙古马背臀部皮下脂肪组织,通过Ⅰ型胶原酶消化、离心等步骤分离培养脂肪组织来源的间充质干细胞(Adipose tissue-derived mesenchymal stem cells,ADSCs),经过原代培养和传代培养,分别加入成脂诱导剂和成骨诱导剂培养,采用倒置显微镜观察诱导后的细胞形态变化,并通过油红O染色和茜素红染色法对其脂肪细胞和成骨细胞表型进行鉴定.结果显示:ADSCs呈成纤维细胞样贴壁生长,其经成脂、成骨诱导培养2周后形态、体积发生明显改变.经油红O染色,细胞质内出现橙红色脂滴;茜素红染色表明聚集的细胞团中央能形成钙化结节.说明马ADSCs经体外诱导培养后可向脂肪细胞和成骨细胞分化,并具有明显的成脂和成骨表型,表明ADSCs具有多向分化潜能.

To investigate the multilineage differentiation capacity of mesenchymal stem cells isolated and cultured from equine adipose tissue,adipose tissue-derived mesenchymal stem cells（ADSCs） were obtained from adipose tissue of Mongolia horse.The cells appeared like fibroblast in the culture medium.Adipose tissue was minced and digested with collagenase type I.The obtained cells were plated and expanded in DMEM /F12 medium.Whereas the passage cells were cultured in adipogenisis medium and stained with Oil Red O for identification.The cells were cultivated in osteoblast-inducing culture medium,and osteoblast phenotype was assayed with Alizarin Red staining.The cells were daily observed under inverted microscope.Results indicated that ADSCs grew as adherent cells,appeared like fibroblast in vitro,stably proliferate and passed.Under the inverted microscope,significant lipid drops were found around the cell nucleus after adipogenisis-inducing cultivation.Alizarin Red staining resulted in the formation of mineralized nods in extracellular matrix.It proved that ADSCs isolated and cultured from equine adipose tissue can be induced to adipogenisis and osteo-inducing,suggesting that the cells have multilineage differentiation.