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

Publication year: 2015
Authors: Sorkio AE1, Vuorimaa-Laukkanen EP2, Hakola HM2, Liang H2, Ujula TA3, Valle-Delgado JJ3, Österberg M3, Yliperttula ML4, Skottman H5
Affiliations:

1BioMediTech, University of Tampere FM5/BMT, 33014 University of Tampere, Finland
2Department of Chemistry and Bioengineering, Tampere University of Technology, Tampere, Finland
3Department of Forest Products Technology, School of Chemical Technology, Aalto University, Aalto, Finland
4Division of Biopharmaceutical Sciences, Centre for Drug Research, Faculty of Pharmacy, University of Helsinki, Helsinki, Finland.
5BioMediTech, University of Tampere, Finland

Published in: Biomaterials, Volume 1, May 2015, Pages 257-269
doi: 10.1016/j.biomaterials.2015.02.005

The environmental cues received by the cells from synthetic substrates in vitro are very different from those they receive in vivo. 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.


MP-SPR keywords: collagen film, Langmuir-Schaefer (LS), layer stability, refractive index, thickness, two media method