Sorkio Anni, Porter Patrick J, Juuti-Uusitalo Kati, Meenan Brian J, Skottman Heli, Burke George A
1 BioMediTech, University of Tampere , Tampere, Finland .
2 Nanotechnology and Integrated Bioengineering Centre (NIBEC), School of Engineering, University of Ulster , Newtownabbey, Northern Ireland .
Tissue Eng Part A. 2015 Sep;21(17-18):2301-14. doi: 10.1089/ten.TEA.2014.0640. Epub 2015 Jun 30.
Human embryonic stem cell-derived retinal pigment epithelial (hESC-RPE) cells are currently undergoing clinical trials to treat retinal degenerative diseases. Transplantation of hESC-RPE cells in conjuction with a supportive biomaterial carrier holds great potential as a future treatment for retinal degeneration. However, there has been no such biodegradable material that could support the growth and maturation of hESC-RPE cells so far. The primary aim of this work was to create a thin porous poly (L-lactide-co-caprolactone) (PLCL) membrane that could promote attachment, proliferation, and maturation of the hESC-RPE cells in serum-free culture conditions. The PLCL membranes were modified by atmospheric pressure plasma processing and coated with collagen IV to enhance cell growth and maturation. Permeability of the membranes was analyzed with an Ussing chamber system. Analysis with scanning electron microscopy, contact angle measurement, atomic force microscopy, and X-ray photoelectron spectroscopy demonstrated that plasma surface treatment augments the surface properties of the membrane, which enhances the binding and conformation of the protein. Cell proliferation assays, reverse transcription-polymerase chain reaction, indirect immunofluoresence staining, trans-epithelial electrical resistance measurements, and in vitro phagocytosis assay clearly demonstrated that the plasma treated PLCL membranes supported the adherence, proliferation, maturation and functionality of hESC-RPE cells in serum-free culture conditions. Here, we report for the first time, how PLCL membranes can be modified with atmospheric pressure plasma processing to enable the formation of a functional hESC-RPE monolayer on a porous biodegradable substrate, which have a potential as a tissue-engineered construct for regenerative retinal repair applications.
人胚胎干细胞来源的视网膜色素上皮（hESC-RPE）细胞目前正在进行治疗视网膜退行性疾病的临床试验。将hESC-RPE细胞与支持性生物材料载体联合移植作为视网膜变性的未来治疗方法具有巨大潜力。然而，迄今为止，尚无能够支持hESC-RPE细胞生长和成熟的可生物降解材料。这项工作的主要目的是制备一种薄的多孔聚（L-丙交酯-共-己内酯）（PLCL）膜，该膜能够在无血清培养条件下促进hESC-RPE细胞的附着、增殖和成熟。通过大气压等离子体处理对PLCL膜进行改性，并涂覆IV型胶原以促进细胞生长和成熟。使用尤斯灌流室系统分析膜的渗透性。通过扫描电子显微镜、接触角测量、原子力显微镜和X射线光电子能谱分析表明，等离子体表面处理增强了膜的表面性质，从而增强了蛋白质的结合和构象。细胞增殖试验、逆转录-聚合酶链反应、间接免疫荧光染色、跨上皮电阻测量和体外吞噬试验清楚地表明，经等离子体处理的PLCL膜在无血清培养条件下支持hESC-RPE细胞的黏附、增殖、成熟和功能。在此，我们首次报道了如何通过大气压等离子体处理对PLCL膜进行改性，以在多孔可生物降解基质上形成功能性hESC-RPE单层，该基质具有作为用于再生视网膜修复应用的组织工程构建体的潜力。
