\r杨 军^{1, 2}，谢敬辉^{1, 2}，高 超^{1, 2}，刘开政^{1, 2}

AuthorsHTML:\r杨 军^{1, 2}，谢敬辉^{1, 2}，高 超^{1, 2}，刘开政^{1, 2}

AuthorsListE:\rYang Jun^{1, 2}，Xie Jinghui^{1, 2}，Gao Chao^{1, 2}，Liu Kaizheng^{1, 2}

AuthorsHTMLE:\rYang Jun^{1, 2}，Xie Jinghui^{1, 2}，Gao Chao^{1, 2}，Liu Kaizheng^{1, 2}

Unit:\r1. 南开大学生命科学学院，天津 300071；
2. 南开大学生物活性材料教育部重点实验室，天津 300071

Unit_EngLish:\r1. College of Life Science，Nankai University，Tianjin 300071，China；
2. Key Laboratory of Bioactive Materials of Ministry of Education，Nankai University，Tianjin 300071，China\r
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Abstract_Chinese:\r为了提高透明质酸水凝胶的生物学功能，本研究利用两步迈克尔加成反应及点击化学反应制备了人血管内皮钙黏素融合蛋白(VE-cad-Fc)功能化的透明质酸水凝胶，并进一步研究了其对人脐带间充质干细胞(hUC-MSCs)的影响．核磁共振光谱检测及凝胶时间检测显示，透明质酸主链丙烯酰肼取代率为16%,，融合蛋白功能化透明质酸以DTT 交联的凝胶化时间约为30,min．ELISA 检测显示水凝胶中化学键合Fc 结构域特异性结合多肽可以显著提高融合蛋白的长期负载稳定性．2D/3D 细胞黏附、增殖与活细胞染色实验结果显示，水凝胶中利用Fc 结构域固定融合蛋白显著改善人脐带间充质干细胞的黏附、增殖，且促进水凝胶中hUC-MSCs 的长期克隆化生长．作为一种细胞培养基质(2D/3D 培养)，这种融合蛋白功能化的水凝胶材料在组织工程、再生医学等领域将有非常广阔的应用前景．\r
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Abstract_English:\rTo improve the biofunction of hyaluronic acid(HA) hydrogel, acryloyl hydrazide hyaluronic acid(HAAC) was first synthesized by Michael addition reaction and click chemistry reaction, and then functionalized by human vascular endothelial cadherin fusion protein(VE-cad-Fc). Subsequently，its impact on the culturing of human
umbilical cord mesenchymal stem cell(hUC-MSCs)was studied. Nuclear magnetic resonance spectroscopy and gelation time detection showed that the substitution rate of acrylation was 16% for HA，and the gelation time of the fusion protein-modified HA-AC upon mixture with DTT crosslinking agent was about 30 min. The quantitative determination of VE-cad-Fc by enzyme-linked immunosorbent assay(ELISA) showed that Fc domain specific binding(Fc-binding) polypeptides chemically bound in the HA-AC can significantly enhance the long-term stability of the fusion protein. The impacts of the hydrogel on hUC-MSCs was further characterized by a live cell staining experiment and the assay of cell adhesion and proliferation in 2D/3D cell culture. The results showed that immobilization of fusion protein in the hydrogel by Fc domain can significantly improve the adhesion，proliferation，and long-term clonal growth of hUCMSCs in the gel. As a cell culture substrate(2D/3D culture)，the fusion protein-functionalized hydrogel can be widely applied in several fields，such as tissue engineering and regenerative medicine.\r
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Keyword_Chinese:水凝胶；透明质酸；人脐带间充质干细胞；细胞黏附；细胞增殖\r

Keywords_English:hydrogel；hyaluronic acid；human umbilical cord mesenchymal stem cell；cell adhesion；cell proliferation\r


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