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原代SD仔鼠心肌成纤维细胞分离与培养方法的改进 被引量:4
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作者 张平 谭延振 +6 位作者 张冰 赵荣 金振晓 易定华 孙阳 易蔚 李香敏 《海南医学》 CAS 2021年第22期2857-2860,共4页
目的探索更好的原代心肌成纤维细胞分离与培养的方法。方法将取出的10只SD仔鼠心室组织置入预冷的PBS中修剪、冲洗后移入无菌玻璃瓶中剪碎;应用胰酶消化液进行初步消化;应用磁力搅拌器搅拌代替滴管吹打混匀Ⅱ型胶原酶消化液与心肌组织... 目的探索更好的原代心肌成纤维细胞分离与培养的方法。方法将取出的10只SD仔鼠心室组织置入预冷的PBS中修剪、冲洗后移入无菌玻璃瓶中剪碎;应用胰酶消化液进行初步消化;应用磁力搅拌器搅拌代替滴管吹打混匀Ⅱ型胶原酶消化液与心肌组织块促进消化顺利进行;应用差速贴壁方法分离成纤维细胞与心肌细胞;应用倒置显微镜观察细胞形态均一性、免疫荧光染色检测Vimentin及CD31表达鉴定心肌成纤维细胞纯度;检测心肌成纤维细胞对TGF-β_(1)诱导的α-平滑肌肌动蛋白(α-SMA)表达变化评估分离的心肌成纤维细胞质量。结果磁力搅拌器搅拌混匀胶原酶消化液与心肌组织块的方法引起的细胞损伤小,消化充分,分离细胞纯度高(Vimentin+/CD-31-,P2代成纤维细胞纯度可达95%以上),细胞活力好,贴壁时间短(约20 min已开始贴壁,90 min贴壁完全),分离所的心肌成纤维细胞激活程度低(α-SMA表达水平低),对TGF-β_(1)诱导刺激反应好(α-SMA表达水平急剧升高)。结论磁力搅拌器方法可成功分离建立高质量的心肌成纤维细胞系。 展开更多
关键词 SD仔鼠 心肌成纤维细胞 细胞培养 磁力搅拌器 差速贴壁方法
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Application of modified polyethylene glycol hydrogels in the construction of tissue engineered heart valve
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作者 OUYANG Hui ZHAO Rong +8 位作者 ZHANG Jin-bao LIU Yang ZHENG Qi-jun YANG Jian GU Chun-hu WEI Xu-feng CHEN Chang-sheng yi ding-hua LIU Wei-yong 《Journal of Life Sciences》 2008年第5期1-9,共9页
To enhance the adhesion of seeding-cells to the biomaterial scaffolds, the PEG-hydrogels were modified. Porcine aortic valves were decellularized with Triton X-100 and trypsin. The cells were encapsulated into the PEG... To enhance the adhesion of seeding-cells to the biomaterial scaffolds, the PEG-hydrogels were modified. Porcine aortic valves were decellularized with Triton X-100 and trypsin. The cells were encapsulated into the PEG-hydrogels to complete the process of the cells attaching to the acellular porcine aortic valves. Herein, the autologous mesenchymal stem cells (MSCs) of goats were selected as the seeding-cells and the tendency of MSCs toward differentiation was observed when the single semilunar TEHV had been implanted into their abdominal aortas. Furthermore, VEGF, TGF-β1, and the cell adhesive peptide motif RGD were incorporated. Light and electron microscopy observations were performed. Analysis of modified PEG-hydrogels TEHV's (PEG-TEHV) tensile strength, and the ratio of reendothelial and mural thrombosis revealed much better improvement than the naked acellular porcine aortic valve (NAPAV). The data illustrated the critical importance of MSC differentiation into endothelial and myofibroblast for remodeling into native tissue. Our results indicate that it is feasible to reconstruct TEHV efficiently by combining modified PEG-hydrogels with acellular biomaterial scaffold andautologous MSCs cells. 展开更多
关键词 tissue engineering BIOMATERIALS DECELLULARIZATION polyethylene glycol hydrogel heart valves
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