title: Biomimetic collagen I and IV double layer Langmuir-Schaefer films as microenvironment for human pluripotent stem cell derived retinal pigment epithelial cells.
abstract: 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.
authors.affiliation: BioMediTech, University of Tampere FM5/BMT, 33014 University of Tampere, Finland. Electronic address: anni.sorkio@uta.fi.
authors.forename: Anni E
authors.initials: AE
authors.lastname: Sorkio
authors.affiliation: Department of Chemistry and Bioengineering, Tampere University of Technology, Korkeakoulunkatu 8, 33720 Tampere, Finland.
authors.forename: Elina P
authors.initials: EP
authors.lastname: Vuorimaa-Laukkanen
authors.affiliation: Department of Chemistry and Bioengineering, Tampere University of Technology, Korkeakoulunkatu 8, 33720 Tampere, Finland.
authors.forename: Hanna M
authors.initials: HM
authors.lastname: Hakola
authors.affiliation: Department of Chemistry and Bioengineering, Tampere University of Technology, Korkeakoulunkatu 8, 33720 Tampere, Finland.
authors.forename: Huamin
authors.initials: H
authors.lastname: Liang
authors.affiliation: Department of Forest Products Technology, School of Chemical Technology, Aalto University, P.O. Box 16300, 00076 Aalto, Finland.
authors.forename: Tiina A
authors.initials: TA
authors.lastname: Ujula
authors.affiliation: Department of Forest Products Technology, School of Chemical Technology, Aalto University, P.O. Box 16300, 00076 Aalto, Finland.
authors.forename: Juan José
authors.initials: JJ
authors.lastname: Valle-Delgado
authors.affiliation: Department of Forest Products Technology, School of Chemical Technology, Aalto University, P.O. Box 16300, 00076 Aalto, Finland.
authors.forename: Monika
authors.initials: M
authors.lastname: Österberg
authors.affiliation: Division of Biopharmaceutical Sciences, Centre for Drug Research, Faculty of Pharmacy, University of Helsinki, Viikinkaari 5, 00790 Helsinki, Finland.
authors.forename: Marjo L
authors.initials: ML
authors.lastname: Yliperttula
authors.affiliation: BioMediTech, University of Tampere FM5/BMT, 33014 University of Tampere, Finland.
authors.forename: Heli
authors.initials: H
authors.lastname: Skottman
mesh_terms: D040761:Biomimetic Materials / Q000494:pharmacology*; D049109:Cell Proliferation / Q000187:drug effects; D060833:Cellular Microenvironment / Q000187:drug effects*; D024042:Collagen Type I / Q000494:pharmacology*; D024141:Collagen Type IV / Q000494:pharmacology*; D006801:Humans; D018625:Microscopy, Atomic Force; D049329:Nanostructures / Q000737:chemistry; D039904:Pluripotent Stem Cells / Q000166:cytology* / Q000187:drug effects; D011312:Pressure; D055213:Retinal Pigment Epithelium / Q000166:cytology* / Q000187:drug effects; D020349:Surface Plasmon Resonance; D013499:Surface Properties; D013696:Temperature
chemical_list: D024042:Collagen Type I; D024141:Collagen Type IV
keywords: Biomimetic material; Collagen structure; Human embryonic stem cell; Langmuir Blodgett film; Retina; Retinal pigment epithelial cell