| 摘要: | 每年有數百萬人因意外、缺陷和疾病遭受器官、組織損害。幹細胞，如人胚胎胎幹細胞（hESCs）和人誘導多能性幹細胞（hiPSCs），具相當潛力應用於轉譯醫學及藥物篩選。使用人類多能性幹細胞細胞進行幹細胞治療臨床試驗有四主要疾病：（1）黃斑部病變 [老年性黃斑部病變與斯塔加特氏病]，（2）急性心肌梗塞，（3）糖尿病（4）脊髓損傷。尤在數據庫分析中發現目前針對hPSCs有效分化成視網膜色素上皮細胞之方法仍不明確。因此開發新穎生醫材料分子，具有最適物理和生物設計相當重要。本研究擬將開發最適細胞培養生醫材料，於無異種條件下將hPSCs分化為RPE。本研究擬（i）將細胞外基質（ECM）及其衍生的奈米片段固定在細胞培養生醫材料上以培養及分化hPSCs（生物因子）和（ii）hPSCs分化為RPE的最適生醫材料軟硬度篩選 (物理因子)。本研究之新創性在幹細胞培養和分化的生醫材料開發係結合物理因子與生物因子。擬以乙烯醇與衣康酸 (PVA-IA)共聚合水凝膠作為基礎生醫材料，以控制交聯時間製備具不同軟硬度PVA-IA之生物不反應性水凝膠。在本研究中，數種ECM蛋白和ECM衍生的寡肽被接枝在具有不同軟硬度之PVA-IA水凝膠用於培養hESC和hiPSC，並以本研究中開發之分化方法，分化成RPE。我們將評估RPE分化的效率，並篩選用於將hPSC特異性分化為RPE的最佳細胞培養生醫材料。 ;Millions of people suffer from organ and tissue loss and damage every year from accidents, birth defects, and diseases. Stem cells, such as human embryonic stem (hES) cells and human induced pluripotent stem (hiPS) cells, are attractive prospects for translational medicine and drug discovery. Four main diseases are targeted for the clinical trials for stem cell therapy using hPS (human pluripotent stem) cells (mainly hES cells): (1) macular degeneration [age-related macular degeneration (AMD) and Stargardt’s macular dystrophy], (2) acute myocardial infarction (MI), (3) diabetes, and (4) spinal cord injury. Especially, efficient differentiation method of hPS cells into retinal pigment epithelium (RPE, for treatment of macular degeneration) has not yet investigated from our database research. Especially, it is unclear what kinds of molecular, physical, and biological design of cell culture biomaterials are preferable for differentiation of hPS cells into RPE. Therefore, the development of sophisticated biomaterials, which are optimized for the differentiation of hPS cells into RPE is important for future stem cell therapies. In this study, we are going to develop optimal cell culture biomaterials for differentiation of hPS cells into RPE in xeno-free and chemically-defined conditions. We will investigate (i) what kind of extracellular matrix (ECM) and ECM-derived nanosegments should be immobilized on cell culture biomaterials for culture and differentiation of hPS cells (biological cues) and (ii) which elasticity is the optimal for hPS cells to differentiate into RPE. Our originality of the biomaterial development for stem cell culture and differentiation is the combination of physical cues (elasticity of biomaterials) and biological cues (selection of ECM or ECM-derived oligopeptide). We are going to develop cell culture biomaterials grafted with ECM and ECM-derived oligopeptides having optimal elasticity, which are suitable for clinical usage in future. Poly(vinylalcohol-co-itaconic acid), PVA-IA, hydrogels are selected as the base biomaterials because PVA-IA hydrogels having different elasticity (i.e., different water content) can be prepared by controlling the crosslinking amount and PVA-IA hydrogels are known to be bioinert. Several ECM proteins and ECM-derived oligopeptides are grafted on the PVA-IA hydrogels having different elasticity in this project. hES cells and hiPS cells are cultured and are differentiated into RPE on the PVA-IA hydrogels grafted with ECM and ECM-derived oligopeptides using differentiation protocols developed in this study. We evaluate the efficiency of RPE differentiation from the data and discuss the optimal cell culture biomaterials for specific differentiation of hPS cells into RPE. |
