TY - JOUR
T1 - Biomimetic collagen I and IV double layer Langmuir-Schaefer films as microenvironment for human pluripotent stem cell derived retinal pigment epithelial cells
AU - Sorkio, Anni E.
AU - Vuorimaa-Laukkanen, Elina P.
AU - Hakola, Hanna M.
AU - Liang, Huamin
AU - Ujula, Tiina A.
AU - Valle-Delgado, Juan Jose
AU - Osterberg, Monika
AU - Yliperttula, Marjo L.
AU - Skottman, Heli
PY - 2015/5
Y1 - 2015/5
N2 - 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 LangmuireSchaefer (LS) deposition, a variant of LangmuireBlodgett 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.
AB - 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 LangmuireSchaefer (LS) deposition, a variant of LangmuireBlodgett 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.
KW - Biomimetic material
KW - Collagen structure
KW - Langmuir Blodgett film
KW - Retinal pigment epithelial cell
KW - Retina
KW - Human embryonic stem cell
U2 - 10.1016/j.biomaterials.2015.02.005
DO - 10.1016/j.biomaterials.2015.02.005
M3 - Article
SN - 0142-9612
VL - 51
SP - 257
EP - 269
JO - Biomaterials
JF - Biomaterials
ER -