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用于间充质干细胞培养的E-钙黏蛋白融合蛋白表面修饰

Surface modification with E-cadherin fusion protein for mesenchymal stem cell culture.

作者信息

Zhang Yan, Mao Hongli, Qian Mengyuan, Hu Feifei, Cao Lei, Xu Ke, Shuai Qizhi, Gao Chao, Lang Ren, Akaike Toshihiro, Yang Jun

机构信息

The Key Laboratory of Bioactive Materials, Ministry of Education, College of Life Science, Nankai University, Tianjin, 300071, China.

出版信息

J Mater Chem B. 2016 Jun 28;4(24):4267-4277. doi: 10.1039/c6tb00765a. Epub 2016 May 31.

Abstract

To effectively expand human mesenchymal stem cells (hMSCs) in vitro without affecting their innate biological properties, a fusion protein (hE-cad-Fc) consisting of a human E-cadherin extracellular domain and an immunoglobulin G Fc region was fabricated and used as a biomimetic matrix for MSC culture surface modification. The results showed that cells cultured on hE-cad-Fc-modified polystyrene surfaces exhibited improved proliferation and paracrine functions compared with cells cultured on unmodified and collagen-modified polystyrene surfaces. Meanwhile, surfaces modified with hE-cad-Fc effectively inhibited cell apoptosis even under the serum deprivation conditions. Additionally, the hE-cad-Fc not only up-regulated the expression of β-catenin in MSCs and stimulated the cellular membrane complex of E-cadherin/β-catenin, but also effectively activated the intracellular signals such as EGFR, AKT and ERK phosphorylation. Therefore, hE-cad-Fc appeared to be a promising candidate for biological surface modification and stem cell culture.

摘要

为了在不影响人骨髓间充质干细胞(hMSCs)固有生物学特性的情况下有效地在体外扩增它们,制备了一种由人E-钙黏蛋白胞外域和免疫球蛋白G Fc区组成的融合蛋白(hE-cad-Fc),并将其用作用于MSC培养表面修饰的仿生基质。结果表明,与在未修饰和胶原修饰的聚苯乙烯表面上培养的细胞相比,在hE-cad-Fc修饰的聚苯乙烯表面上培养的细胞表现出更好的增殖和旁分泌功能。同时,即使在血清剥夺条件下,用hE-cad-Fc修饰的表面也能有效抑制细胞凋亡。此外,hE-cad-Fc不仅上调了MSCs中β-连环蛋白的表达并刺激了E-钙黏蛋白/β-连环蛋白的细胞膜复合物,还有效激活了细胞内信号,如EGFR、AKT和ERK磷酸化。因此,hE-cad-Fc似乎是生物表面修饰和干细胞培养的一个有前途的候选物。

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