TY - JOUR

T1 - Culture surface protein coatings affect the barrier properties and calcium signalling of hESC-RPE

AU - Viheriälä, Taina

AU - Sorvari, Juhana

AU - Ihalainen, Teemu O.

AU - Mörö, Anni

AU - Grönroos, Pyry

AU - Schlie-Wolter, Sabrina

AU - Chichkov, Boris

AU - Skottman, Heli

AU - Nymark, Soile

AU - Ilmarinen, Tanja

N1 - Funding Information: The authors thank biomedical laboratory technicians Outi Melin, Hanna Pekkanen, and Emma Vikstedt for their technical assistance and contributions to cell culture. The authors acknowledge the Biocenter Finland and Tampere Imaging Facility for their services. This study was supported by the Academy of Finland grant numbers 287287, 304909, 308315, 319257, 323508), the Instrumentarium Science Foundation (170050, 180038), the Finnish Cultural Foundation (00171144, 00181174) and the Emil Aaltonen Foundation.

PY - 2021/1/13

Y1 - 2021/1/13

N2 - Human pluripotent stem cell-derived retinal pigment epithelium (RPE) transplantation is currently under evaluation as treatment for macular degeneration. For therapeutic applications, cryostorage during cell production is typically needed with potential consequences to cell functionality. We have previously shown that the culture substrate affects human embryonic stem cell-derived RPE (hESC-RPE) properties in fresh cultures. Here, we aimed to further identify the role of RPE basement membrane proteins type IV collagen (Col-IV), laminin (LN), and nidogen-1 in the maturation and functionality of hESC-RPE after cryopreservation. In addition to cell attachment and morphology, transepithelial electrical resistance, expression of key RPE proteins, phagocytosis capacity and Ca2+ signalling were analysed. After cryostorage, attachment of hESC-RPE on culture surfaces coated with Col-IV alone was poor. Combining Col-IV and LN with or without nidogen-1 significantly improved cell attachment and barrier properties of the epithelium. Furthermore, functional homogeneity of the hESC-RPE monolayer was enhanced in the presence of nidogen-1. Our results suggest that the choice of coating proteins for the cell culture may have implications to the functional properties of these cells after cryostorage cell banking.

AB - Human pluripotent stem cell-derived retinal pigment epithelium (RPE) transplantation is currently under evaluation as treatment for macular degeneration. For therapeutic applications, cryostorage during cell production is typically needed with potential consequences to cell functionality. We have previously shown that the culture substrate affects human embryonic stem cell-derived RPE (hESC-RPE) properties in fresh cultures. Here, we aimed to further identify the role of RPE basement membrane proteins type IV collagen (Col-IV), laminin (LN), and nidogen-1 in the maturation and functionality of hESC-RPE after cryopreservation. In addition to cell attachment and morphology, transepithelial electrical resistance, expression of key RPE proteins, phagocytosis capacity and Ca2+ signalling were analysed. After cryostorage, attachment of hESC-RPE on culture surfaces coated with Col-IV alone was poor. Combining Col-IV and LN with or without nidogen-1 significantly improved cell attachment and barrier properties of the epithelium. Furthermore, functional homogeneity of the hESC-RPE monolayer was enhanced in the presence of nidogen-1. Our results suggest that the choice of coating proteins for the cell culture may have implications to the functional properties of these cells after cryostorage cell banking.

UR - http://www.scopus.com/inward/record.url?scp=85099354546&partnerID=8YFLogxK

U2 - 10.1038/s41598-020-79638-8

DO - 10.1038/s41598-020-79638-8

M3 - Article

C2 - 33441679

AN - SCOPUS:85099354546

VL - 11

JO - Scientific reports

JF - Scientific reports

SN - 2045-2322

M1 - 933

ER -