中国组织工程研究Issue(52)：8474-8479,6.DOI:10.3969/j.issn.2095-4344
胶原补片上生长因子释放及对人骨髓干细胞增殖与分化的影响
Release and effect of growth factors conjugated into collagen patches on human bone marrow mesenchymal stem cell proliferation and differentiation
摘要
Abstract
BACKGROUND:Preclinical trials in animals have demonstrated myocardial cel transplantation can improve heart function, but it is hindered by the low retention of transplanted cels. Therefore, it becomes a common strategy to generate a proper cel platform to promote cel retention and vasculature formation. <br> OBJECTIVE:To evaluate the cytokine release of a novel cytokine-conjugated colagen patch prepared with 1-ethyl-3-[3-dimethylaminopropyl] carbodimide hydrochloride (EDC) and its physiological effects on proliferation and differentiation of seeded human bone marrow mesenchymal stem cels. <br> METHODS:After activating and crosslinking by EDC, the control patches (EDC-patch) were preserved in PBS and the experimental patches (GF-patch) were immersed into PBS containing vascular endothelial growth factor and basic fibroblast growth factor for cytokine conjugation. Colagen patches without EDC treatment underwent same process as internal control. ELISA was performed to detect the content of vascular endothelial growth factor and basic fibroblast growth factor in the supernatant of GF-patch at 1, 3, 7 days, 2, 3, 4 weeks after patch preparation. The 0.5×106 human bone marrow mesenchymal stem cels were evenly seeded on both kinds of patches and the cel proliferation was identified by hematoxylin-eosin staining folowed by cel counting, MTT assay or BrdU staining test. Accordingly, cel differentiation was ilustrated by RT-PCR for colagen I and colagen III expression, and immunofluorescent staining for SMA in seeding cels. <br> RESULTS AND CONCLUSION: 42.4% vascular endothelial growth factors and 24.5% basic fibroblast growth factors were successfuly conjugated into colagen patches and they both exhibited the constant, controled-release mode during the 4-week observation period. By contrast, the patch without EDC treatment just showed the physical bonding of both cytokines. In comparison with EDC-patch, GF-patch presented the capability to induce cel proliferation while retard cel differentiation.关键词
生物材料/缓释材料/心脏补片/血管内皮生长因子/碱性成纤维细胞生长因子/细胞移植/载体/碳化二亚胺法/生长因子/缓释/血管形成/国家自然科学基金Key words
Subject headings:colagen/carbodimides/vascular endothelial growth factors/fibroblast growth factor 2分类
医药卫生引用本文复制引用
啜俊波,李仁科,康凯,孙露,曲辉,蔡俊,陈克功,谢宝栋,蒋树林,田海..胶原补片上生长因子释放及对人骨髓干细胞增殖与分化的影响[J].中国组织工程研究,2014,(52):8474-8479,6.基金项目
黑龙江省卫生厅课题资助项目(2013053)；国家自然科学基金(81471805)；中国博士后科研基金(2014M551272)；黑龙江省教育厅科研课题(12541434)Funding：the Project of Heilongjiang Health Bureau, No.2013053 （2013053）
the National Natural Science Foundation of China, No.81471805 （）
the China Postdoctoral Science Foundation, No.2014M551272 （）
the Project of Heilongjiang Education Bureau, No.12541434 （）
