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Biomimetic collagen I and IV double layer Langmuir-Schaefer films asmicroenvironment for human pluripotent stem cell derived retinal pigment epithelial cells

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Details

Original languageEnglish
Pages (from-to)257-269
Number of pages13
JournalBiomaterials
Volume51
DOIs
Publication statusPublished - 1 May 2015
Publication typeA1 Journal article-refereed

Abstract

The environmental cues received by the cells from synthetic substrates invitro are very different from those they receive invivo. In this study, we applied the Langmuir-Schaefer (LS) deposition, a variant of Langmuir-Blodgett technique, to fabricate a biomimetic microenvironment mimicking the structure and organization of native Bruch's membrane for the production of the functional human embryonic stem cell derived retinal pigment epithelial (hESC-RPE) cells. Surface pressure-area isotherms were measured simultaneously with Brewster angle microscopy to investigate the self-assembly of human collagens type I and IV on air-subphase interface. Furthermore, the structure of the prepared collagen LS films was characterized with scanning electron microscopy, atomic force microscopy, surface plasmon resonance measurements and immunofluorescent staining. The integrity of hESC-RPE on double layer LS films was investigated by measuring transepithelial resistance and permeability of small molecular weight substance. Maturation and functionality of hESC-RPE cells on double layer collagen LS films was further assessed by RPE-specific gene and protein expression, growth factor secretion, and phagocytic activity. Here, we demonstrated that the prepared collagen LS films have layered structure with oriented fibers corresponding to architecture of the uppermost layers of Bruch's membrane and result in increased barrier properties and functionality of hESC-RPE cells as compared to the commonly used dip-coated controls.

Keywords

  • Biomimetic material, Collagen structure, Human embryonic stem cell, Langmuir Blodgett film, Retina, Retinal pigment epithelial cell

Publication forum classification

Field of science, Statistics Finland