重组人干扰素β1a对人骨髓间充质干细胞成软骨细胞定向分化的影响

Effects of recombinant human interferon β1a on the chondrogenic differentiation of human bone marrow mesenchymal stem cell

摘要:

重组人干扰素β (rhIFN-β) 是通过基因表达的一种糖蛋白，具有抗病毒、抗肿瘤和免疫调节作用，虽有研究表明其他亚型干扰素如干扰素γ对细胞增殖和分化有一定的作用，但rhIFN-β对人骨髓间充质干细胞 (hMSCs) 诱导分化的影响尤其是成软骨细胞定向分化的研究尚少。文中采用成球法研究在常规TGF-β3诱导分化培养基中添加重组IFN-β1a后对诱导hMSCs分化成软骨球的影响。hMSCs经诱导分化后，收集软骨球，通过定量检测糖胺多糖 (GAG) 含量、软骨球形态测定、Alcian Blue 组织染色、Real-time PCR和Western blotting检测Sox9和Collangen Ⅱ的表达。结果显示，在常规TGF-β3诱导分化培养基中添加100 ng/mL IFN-β1a能显著提高GAG含量，增大软骨球尺寸，促进聚集蛋白聚糖形成，上调Sox9和Collangen Ⅱ的表达。研究结果表明，重组人IFN-β1a能够与TGF-β3联合作用促进hMSCs成软骨细胞定向分化。

Abstract:

Recombinant human interferon beta (rhIFN-β) is a glycoprotein produced by genetically engineered cells and has anti-virus, anti-tumor and immunoregulation functions. Although studies have shown that other subtypes of IFN such as IFN-γ affects cell proliferation and differentiation to some extent, the effect of rhIFN-β on chondrogenic differentiation of human bone marrow mesenchymal stem cells (hMSCs) is less known. In this study we studied the effect of rhIFN-β on the chondrogenic differentiation of hMSCs by inducing hMSCs into cartilage pellet via adding IFN-β1a into regular TGF-β3 chondrogenic differentiation medium. We collected the induced pellets and then detected GAG content, assessed pellets size, observed agreecan using alcian blue staining, and analyzed the expression of Sox and CollangenⅡusing real-time PCR and Western blotting. Addition of 100 ng/mL IFN-β1a to regular TGF-β3 chondrogenic differentiation medium could improve the concentration of GAG, increase the size of pellets, promote the formation of aggrecan and up-regulate the expression of CollangenII and Sox9. IFN-β1a combined with TGF-β3 could promote chondrogenic differentiation of hMSCs.