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Collagen vitrigels with low‐fibril density enhance human embryonic stem cell‐derived retinal pigment epithelial cell maturation
Authors:Xiaokun Wang  Julien Maruotti  Shoumyo Majumdar  Jose Roman  Hai‐Quan Mao  Donald J. Zack  Jennifer H. Elisseeff
Affiliation:1. Wilmer Eye Institute, School of Medicine, Johns Hopkins University, Baltimore, MD, USA;2. Phenocell SAS, Evry, France;3. Department of Materials Science and Engineering, Johns Hopkins University, Baltimore, MD, USA;4. Department of Biomedical Engineering, Johns Hopkins University, Baltimore, MD, USA
Abstract:Structural and biochemical cues of extracellular matrix can substantially influence the differentiation and maturation of cultured retinal pigment epithelial (RPE) cells. In this study, thin collagen vitrigels were engineered to create collagen nanofibrillar structures of different fibril densities in an effort to evaluate the maturation of human embryonic stem cell–derived retinal pigment epithelial (hESC‐RPE) cells. The ultrastructure of the different collagen vitrigels was characterized by transmission electron microscopy, and the mechanical properties were evaluated by tensile testing. The pigmentation and polarization of cells, in addition to key RPE marker gene and protein expression levels, were analyzed to determine the differentiation of hESCs on the gels. The hESC‐RPE differentiation was most significant in collagen vitrigels with low fibril density with mature collagen fibrils with diameter of around 60 nm and Young's modulus of 2.41 ± 0.13 MPa. This study provides insight into the influence of collagen nanofibrillar structures on hESC‐RPE maturation and presents a potential bioengineered substratum for hESC‐RPE for future preclinical and clinical applications.
Keywords:cell maturation  collagen vitrigel  epithelial cells  fibril density  retina  stem cells
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