人脂肪源性干细胞多潜能分化特性

The proliferation and differentiation of human adipose-derived stem cells

目的原代分离培养人脂肪干细胞(ADSCs),探讨细胞增殖及多向分化,间充质干细胞相关特性。方法取人吸脂术的脂肪组织通过酶消化法分离培养ADSCs,观察细胞形态,MTT法检测细胞增殖活性,流式细胞术检测细胞周期和表面抗原的表达,用茜素红、甲苯胺蓝和油红O染色鉴定向骨、软骨及脂肪诱导分化。结果人ADSCs呈长梭形、旋涡状生长,传至第15代的ADSCs仍具较强的增殖能力,ADSCs具有干细胞特性,高表达标记CD90和CD44的间充质干细胞,不表达内皮细胞标记CD31,而CD49d低表达,CD106阴性。成骨诱导后可见钙结节形成并被茜素红染成紫红色,软骨诱导后分泌大量蛋白多糖,被甲苯胺蓝染成蓝色,成脂诱导后细胞内可见大量脂滴被油红O染色红色。结论人脂肪干细胞可分化成间充质干细胞,具有稳定增殖和成骨、成软骨和成脂等多向分化潜能。

Objective The human adipose derived stem cells（ADSCs） were isolated and cultured to explore their multipotent differentiation ability. Methods ADSCs were isolated and cultured by enzymatic digestion from fat tissue of human liposuetion aspirates. The cell morphology was observed by phase contrast microscope, the cell viability was evaluated with MTT, the cell cycle and surface marker profiles were detected by flow eytometry. The osteogenie, ehondrogenie and adipogenic differentiations were identified with Alizarin red and toluidine blue, Oil Red O staining respectively. Results The ADSCs showed a vortex growth with fibroblasts-like shape, with strong proliferation activity and characteristics of stem cells when reached passage 15. The positive expressions of mesenehymal stem cells markers CD90 and CD44, negative expression of endothelial cell mark CD31 were observed in ADSCs by flow cytometry, with lower expression of CD49d and negative expression of CD106. Calcium nodules were observed after osteogenic induction and Alizarin red staining was positive, and proteoglycan was secreted after chondrogenic induction and toluidine blue staining was positive. Oil Red O staining of ADSCs after adipogenic induction demonstrated numerous intracellular lipid droplets. Conclusion ADSCs with strong proliferative ability could be differentiated into mesenchymal stem cells, osseous cells, cartilage cells and adipose cells.