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Real-time monitoring of human Schwann cells on heparin-collagen coatings reveals enhanced adhesion and growth factor response.
Journal of Materials Chemistry B ( IF 6.1 ) Pub Date : 2020-08-25 , DOI: 10.1039/d0tb01454k Luis Pinzon-Herrera 1 , Janet Mendez-Vega , Adriana Mulero-Russe , David A Castilla-Casadiego , Jorge Almodovar
Journal of Materials Chemistry B ( IF 6.1 ) Pub Date : 2020-08-25 , DOI: 10.1039/d0tb01454k Luis Pinzon-Herrera 1 , Janet Mendez-Vega , Adriana Mulero-Russe , David A Castilla-Casadiego , Jorge Almodovar
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In this work, we evaluate the enhancing effect of six bilayers of heparin/collagen (HEP/COL)6 layer-by-layer coatings on human Schwann cell (hSCs) adhesion and proliferation in the presence or absence of nerve growth factor (NGF). hSCs behavior and in vitro bioactivity were studied during six days of culture using end-point viability and proliferation assays as well as an impedance-based real-time monitoring system. An end-point viability assay revealed that hSCs cultured on the (HEP/COL)6 coatings increased their growth by more than 230% compared to controls. However, an EdU proliferation assay revealed that the proliferation rate of hSCs in all conditions were similar, with 45% of cells proliferating after 18 hours of incubation. Fluorescence microscopy revealed that hSCs spreading was similar between the tissue culture plastic control and the (HEP/COL)6. The presence of NGF in solution resulted in cells with a larger spread area. Real-time monitoring of hSCs seeded on (HEP/COL)6 with and without NGF reveals that initial cell adhesion is improved by the presence of the (HEP/COL)6 coatings, and it is further improved by the presence of NGF. Our results suggest that (HEP/COL)6 coatings enhance Schwann cell behavior and response to NGF. This simple modification could be applied to current nerve regeneration strategies to improve the repair of damaged nerve.
中文翻译：
肝素-胶原蛋白涂层上的人雪旺氏细胞的实时监控显示增强的粘附和生长因子反应。
在这项工作中，我们评估在存在或不存在神经生长因子（NGF）的情况下，六层肝素/胶原蛋白（HEP / COL）6层涂层对人雪旺细胞（hSCs）粘附和增殖的增强作用。在六天的培养过程中，使用终点活力和增殖测定以及基于阻抗的实时监测系统研究了hSCs的行为和体外生物活性。终点生存力分析表明，在（HEP / COL）6上培养的hSC与对照组相比，涂料的生长速度提高了230％以上。但是，EdU增殖试验表明，在所有条件下，hSC的增殖速率均相似，孵育18小时后，有45％的细胞增殖。荧光显微镜显示，hSCs在组织培养塑料对照和（HEP / COL）6之间扩散相似。溶液中NGF的存在导致细胞具有更大的扩散面积。实时监测在有和没有NGF的情况下接种在（HEP / COL）6上的hSCs显示，通过（HEP / COL）6涂层的存在可以改善初始细胞粘附，而通过存在NGF可以进一步改善。我们的结果表明（HEP / COL）6涂层可增强雪旺氏细胞的行为和对NGF的反应。这种简单的修改可以应用于当前的神经再生策略，以改善受损神经的修复。
更新日期：2020-10-07
中文翻译：
肝素-胶原蛋白涂层上的人雪旺氏细胞的实时监控显示增强的粘附和生长因子反应。
在这项工作中，我们评估在存在或不存在神经生长因子（NGF）的情况下，六层肝素/胶原蛋白（HEP / COL）6层涂层对人雪旺细胞（hSCs）粘附和增殖的增强作用。在六天的培养过程中，使用终点活力和增殖测定以及基于阻抗的实时监测系统研究了hSCs的行为和体外生物活性。终点生存力分析表明，在（HEP / COL）6上培养的hSC与对照组相比，涂料的生长速度提高了230％以上。但是，EdU增殖试验表明，在所有条件下，hSC的增殖速率均相似，孵育18小时后，有45％的细胞增殖。荧光显微镜显示，hSCs在组织培养塑料对照和（HEP / COL）6之间扩散相似。溶液中NGF的存在导致细胞具有更大的扩散面积。实时监测在有和没有NGF的情况下接种在（HEP / COL）6上的hSCs显示，通过（HEP / COL）6涂层的存在可以改善初始细胞粘附，而通过存在NGF可以进一步改善。我们的结果表明（HEP / COL）6涂层可增强雪旺氏细胞的行为和对NGF的反应。这种简单的修改可以应用于当前的神经再生策略，以改善受损神经的修复。
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