摘要
目的分离培养人脂肪干细胞(hADSCs),用不同浓度GDF-5进行诱导,观察细胞形态变化及检测相关蛋白的表达,评价其是否具有软骨样细胞的特性。方法取人抽脂术后的无菌脂肪组织,酶消化法分离培养hADSCs,观察细胞形态,MTT法检测细胞增殖能力。用含0、50、100ng/mlGDF-5的软骨诱导液培养,观察形态变化,检测Ⅱ型胶原及GAG的表达。结果 hADSCs呈长梭形,P10以后仍具有较强的增殖能力。经GDF-5诱导的hADSCs逐渐变为圆形、多角形,部分细胞聚集生长,形成软骨样结节;诱导28d的细胞Ⅱ型胶原表达阳性,甲苯胺蓝将GAG染成蓝色,但100ng/mlGDF-5诱导组阳性表达明显高于50ng/ml诱导组。结论 hADSCs在100ng/mlGDF-5的诱导下可分化为软骨样细胞。
Objective We isolated and cultured human adipose derived stem cells which was induced and differentiated into cartilage-like cells by different concentrations of GDF-5.The morphology and characteristics of induced cells was detected,in order that we can explore more reliable induced method.Methods From the liposuction aspirates,hADSCs was isolated by enzymatic digestion,and the morphology of the cultured cells was observed.The cell viability was evaluated with MTT and cell growth curve was generated.hADSCs was induced and differentiated into cartilage-like cells by different concentrations of GDF-5 （0,50,100 ng/ml）.The morphology was observed.The expression of collagen Ⅱwas assessed by immunocytochemically.The glycosaminoglycan was detected by toluidine blue stain.Results The hADSCs presented long spindle with a fibroblast-like appearance and strong proliferation activity when they spread to passage 10 as shown by MTT.The induced cells in GDF-5 became round and polygon gradually,and formed cartilage-like aggregation.The type Ⅱ collagen was expressed by the cells on induced 21 d.GAG was stained with toluidine blue.But it was significantly higher of 100 ng/ml GDF-5 induced group than that of 50 ng/ml induced group.Conclusion hADSCs possesses stem cell properties and can be differentiated into cartilage-like cells in 100 ng/ml GDF-5.
出处
《中国实用医药》
2012年第1期3-5,共3页
China Practical Medicine
关键词
脂肪干细胞
诱导
分化
软骨样细胞
Human adipose derived stem cells Induced Differentiation Cartilage-like cells
