实时监测人雪旺细胞在肝素-胶原涂层上的情况,发现其黏附性增强,生长因子反应增强。
Real-time monitoring of human Schwann cells on heparin-collagen coatings reveals enhanced adhesion and growth factor response.
机构信息
Ralph E. Martin Department of Chemical Engineering, University of Arkansas, 3202 Bell Engineering Center, Fayetteville, AR 72701, USA.
出版信息
J Mater Chem B. 2020 Oct 14;8(38):8809-8819. doi: 10.1039/d0tb01454k. Epub 2020 Aug 28.
In this work, we evaluate the enhancing effect of six bilayers of heparin/collagen (HEP/COL) 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) 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). The presence of NGF in solution resulted in cells with a larger spread area. Real-time monitoring of hSCs seeded on (HEP/COL) with and without NGF reveals that initial cell adhesion is improved by the presence of the (HEP/COL) coatings, and it is further improved by the presence of NGF. Our results suggest that (HEP/COL) 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.
在这项工作中,我们评估了肝素/胶原蛋白(HEP/COL)六层层层涂层对人雪旺细胞(hSCs)在有或没有神经生长因子(NGF)存在时的黏附和增殖的增强作用。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 在培养 6 天期间的行为和体外生物活性。终点活力测定显示,与对照组相比,培养在(HEP/COL)涂层上的 hSCs 生长增加了 230%以上。然而,EdU 增殖测定显示,所有条件下 hSCs 的增殖率相似,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)之间的铺展情况相似。溶液中 NGF 的存在导致细胞的铺展面积更大。实时监测有和没有 NGF 的(HEP/COL)上接种的 hSCs 显示,(HEP/COL)涂层的存在可改善初始细胞黏附,而 NGF 的存在可进一步改善初始细胞黏附。我们的结果表明,(HEP/COL)涂层可增强雪旺细胞的行为和对 NGF 的反应。这种简单的修饰可以应用于当前的神经再生策略,以改善受损神经的修复。