A novel electro-chemotactic approach to impact the directional migration of transplantable retinal progenitor cells
A novel electro-chemotactic approach to impact the directional migration of transplantable retinal progenitor cells
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DOI：
10.1016/j.exer.2019.06.002
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发表时间：
2019-08-01
影响因子：
3.4
通讯作者：
Vazquez, Maribel
中科院分区：
文献类型：
--
作者：
Mishra, Shawn;Pena, Juan S.;Vazquez, Maribel
光感受器退化是美国和全球视力受损的一个重要原因。细胞替代疗法通过将干细胞移植到视网膜下腔来替代受损的光感受器，在恢复视力方面显示出巨大的前景。然而，通过移植修复视力一直受到限制，在很大程度上是因为能够迁移到受损视网膜组织并与天然光感受器整合的替代细胞数量较少。项目使用外部化学场和外加电场分别诱导替代细胞的趋化性和电趋性，但成效有限。然而，令人惊讶的是，在生物材料和宿主组织中应用组合的电化学趋化场来指导细胞的研究还不够深入。目前的工作检验了在受控微环境中，联合电化学场指导可移植视网膜祖细胞(RPC)迁移的能力。实验使用我们建立的电流-微流控系统(GAL-M-Mu，S)，在有和没有叠加电场的情况下产生了可调的趋化浓度场。结果表明，组合场增加了RPC迁移距离的三倍以上，分别是在任一场中看到的，并且具有更大的方向性来增加梯度。有趣的是，免疫荧光分析显示，总的和/或激活的同源受体的分布没有显著差异，表明配体结合的变化本身并不是测量到的迁移增加的原因。然后进行生物信息学分析，以识别潜在的、参与电趋化的协同机制路径。结果表明，RPC在电化学趋化领域的迁移增加可能是由于细胞黏附蛋白的下调和细胞骨架调节蛋白的上调所致。这些全面的结果指向一种新的移行靶向治疗，它可能显著改善移植结果，并阐明未报道的跨生物机制的协同作用对电化学趋化场的反应。
Photoreceptor degeneration is a significant cause of visual impairment in the United States and globally. Cell replacement therapy shows great promise in restoring vision by transplanting stem-like cells into the sub-retinal space as substitutes for damaged photoreceptors. However, vision repair via transplantation has been limited, in large part, by low numbers of replacement cells able to migrate into damaged retinal tissue and integrate with native photoreceptors. Projects have used external chemical fields and applied electric fields to induce the chemotaxis and electrotaxis of replacement cells, respectively, with limited success. However, the application of combined electro-chemotactic fields in directing cells within biomaterials and host tissue has been surprisingly understudied. The current work examined the ability of combined electro-chemotactic fields to direct the migration of transplantable retinal progenitor cells (RPCs) in controlled microenvironments. Experiments used our established galvano-microfluidic system (Gal-M mu S) to generate tunable chemotactic concentration fields with and without superimposed electric fields. Result illustrate that combination fields increased the distance migrated by RPCs by over three times that seen in either field, individually, and with greater directionality towards increasing gradients. Interestingly, immunofluorescence assays showed no significant differences in the distribution of the total and/or activated cognate receptor of interest, indicating that changes in ligand binding alone were not responsible for the measured increases in migration. Bioinformatics analysis was then performed to identity potential, synergistic mechanistic pathways involved in the electro-chemotaxis measured. Results indicate that increased RPC migration in electro-chemotactic fields may arise from down-regulation of cell adhesion proteins in tandem with up-regulation of cytoskeletal regulation proteins. These comprehensive results point towards a novel migration-targeted treatment that may dramatically improve transplantation outcomes as well as elucidate unreported synergy across biological mechanisms in response to electro-chemotactic fields.