PDF(Pubmed)
Abstract：
Retinal degenerative diseases are a major cause of blindness involving the dysfunction of photoreceptors, retinal pigmented epithelium (RPE), or both. A promising treatment approach involves replacing these cells via surgical transplantation, and previous work has shown that cell delivery scaffolds are vital to ensure sufficient cell survival. Thus, identifying scaffold properties that are conducive to cell viability and maturation (such as suitable material and mechanical properties) is critical to ensuring a successful treatment approach. In this study, we investigated the effect of scaffold stiffness on human RPE attachment, survival, and differentiation, comparing immortalized (ARPE-19) and stem cell-derived RPE (iRPE) cells. Polydimethylsiloxane was used as a model polymer substrate, and varying stiffness (~12 to 800 kPa) was achieved by modulating the cross-link-to-base ratio. Post-attachment changes in gene and protein expression were assessed using qPCR and immunocytochemistry. We found that while ARPE-19 and iRPE exhibited significant differences in morphology and expression of RPE markers, substrate stiffness did not have a substantial impact on cell growth or maturation for either cell type. These results highlight the differences in expression between immortalized and iPSC-derived RPE cells, and also suggest that stiffnesses in this range (~12-800 kPa) may not result in significant differences in RPE growth and maturation, an important consideration in scaffold design.
摘要：
视网膜退行性疾病是导致失明的主要原因，涉及光感受器的功能障碍，视网膜色素上皮（RPE），或者两者兼而有之。一种有前途的治疗方法包括通过手术移植替换这些细胞，和以前的工作已经表明，细胞输送支架是至关重要的，以确保足够的细胞存活。因此，确定有利于细胞活力和成熟的支架特性（例如合适的材料和机械性能）对于确保成功的治疗方法至关重要。在这项研究中,我们研究了支架刚度对人体RPE附着的影响，生存,和差异化，比较永生化（ARPE-19）和干细胞衍生的RPE（iRPE）细胞。聚二甲基硅氧烷用作模型聚合物基材，和变化的刚度（〜12至800kPa）是通过调节交联比实现的。使用qPCR和免疫细胞化学评估基因和蛋白质表达的附着后变化。我们发现，虽然ARPE-19和iRPE在RPE标志物的形态和表达方面表现出显著差异，对于两种细胞类型，底物刚度对细胞生长或成熟没有实质性影响。这些结果突出了永生化和iPSC衍生的RPE细胞之间表达的差异，并且还表明，在此范围内的刚度（〜12-800kPa）可能不会导致RPE生长和成熟的显着差异，脚手架设计中的重要考虑因素。
