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目的:体外分离培养鉴定具有间充质干细胞(mesenchymal stem cell,MSC)特征的人羊水来源细胞,并对其免疫抑制特性进行初步评价。方法:贴壁法体外分离培养人羊水来源细胞,多次传代扩增后,采用形态学观察、流式细胞术分析表型以及定向诱导分化为脂肪样细胞、成骨样细胞等方法进行鉴定后,以细胞计数试剂盒(cell counting kit-8,CCK-8)体外检测人羊水来源间充质干细胞(amniotic fluid derived-mesenchymal stem cells,AFMSC)对ConA刺激的小鼠脾淋巴细胞增殖的影响,并对其时效、量效关系进行分析。结果:具备MSC形态、表型及分化潜能特征的人羊水来源细胞,可于体外显著抑制ConA刺激的小鼠脾淋巴细胞增殖(P<0.05),时效、量效关系显著(P<0.05)。结论:从免疫抑制特性角度证明羊水可成为替代骨髓的MSC新来源。
OBJECTIVE: To isolate and culture in vitro human amniotic fluid derived cells characterized by mesenchymal stem cells (MSCs), and to evaluate their immunosuppressive properties. Methods: Human amniotic fluid derived cells were isolated and cultured in vitro by adherence method. After repeated passages and amplifications, morphological observation, phenotypic analysis by flow cytometry, and differentiated into adipose-like cells and osteoblast-like cells were carried out. To investigate the effect of amniotic fluid derived-mesenchymal stem cells (AFMSC) on the proliferation of ConA-stimulated mouse splenic lymphocytes by cell counting kit-8 (CCK-8) And its aging, the relationship between quantitative and effective analysis. Results: Human amniotic fluid derived cells with the morphological, phenotypic and differentiation potential of MSC could significantly inhibit the proliferation of ConA-stimulated splenic lymphocytes in vitro (P <0.05). The aging and dose-related effects were significant (P <0.05). CONCLUSION: Amniotic fluid can be used as a new source of MSC in place of bone marrow from the perspective of immunosuppressive properties.