Identification of cell surface markers and establishment of monolayer differentiation to retinal pigment epithelial cells.

Autor: Plaza Reyes A; Department of Clinical Sciences, Intervention and Technology, Karolinska Institutet, Stockholm, Sweden.; Ming Wai Lau Center for Reparative Medicine, Stockholm node, Karolinska Institutet, Stockholm, Sweden.; Division of Obstetrics and Gynecology, Karolinska Universitetssjukhuset, Stockholm, Sweden., Petrus-Reurer S; Department of Clinical Sciences, Intervention and Technology, Karolinska Institutet, Stockholm, Sweden.; Ming Wai Lau Center for Reparative Medicine, Stockholm node, Karolinska Institutet, Stockholm, Sweden.; Division of Obstetrics and Gynecology, Karolinska Universitetssjukhuset, Stockholm, Sweden.; Department of Clinical Neuroscience, Division of Eye and Vision, St. Erik Eye Hospital, Karolinska Institutet, Stockholm, Sweden., Padrell Sánchez S; Department of Clinical Sciences, Intervention and Technology, Karolinska Institutet, Stockholm, Sweden.; Ming Wai Lau Center for Reparative Medicine, Stockholm node, Karolinska Institutet, Stockholm, Sweden.; Division of Obstetrics and Gynecology, Karolinska Universitetssjukhuset, Stockholm, Sweden., Kumar P; Department of Clinical Sciences, Intervention and Technology, Karolinska Institutet, Stockholm, Sweden.; Ming Wai Lau Center for Reparative Medicine, Stockholm node, Karolinska Institutet, Stockholm, Sweden.; Division of Obstetrics and Gynecology, Karolinska Universitetssjukhuset, Stockholm, Sweden., Douagi I; Department of Medicine, Center for Hematology and Regenerative Medicine, Karolinska Institutet, Stockholm, Sweden., Bartuma H; Department of Clinical Neuroscience, Division of Eye and Vision, St. Erik Eye Hospital, Karolinska Institutet, Stockholm, Sweden., Aronsson M; Department of Clinical Neuroscience, Division of Eye and Vision, St. Erik Eye Hospital, Karolinska Institutet, Stockholm, Sweden., Westman S; Department of Clinical Neuroscience, Division of Eye and Vision, St. Erik Eye Hospital, Karolinska Institutet, Stockholm, Sweden., Lardner E; Department of Clinical Neuroscience, Division of Eye and Vision, St. Erik Eye Hospital, Karolinska Institutet, Stockholm, Sweden., André H; Department of Clinical Neuroscience, Division of Eye and Vision, St. Erik Eye Hospital, Karolinska Institutet, Stockholm, Sweden., Falk A; Department of Neuroscience, Karolinska Institutet, Stockholm, Sweden., Nandrot EF; INSERM, CNRS, Institut de la Vision, Sorbonne Université, 75012, Paris, France., Kvanta A; Department of Clinical Neuroscience, Division of Eye and Vision, St. Erik Eye Hospital, Karolinska Institutet, Stockholm, Sweden., Lanner F; Department of Clinical Sciences, Intervention and Technology, Karolinska Institutet, Stockholm, Sweden. Fredrik.Lanner@ki.se.; Ming Wai Lau Center for Reparative Medicine, Stockholm node, Karolinska Institutet, Stockholm, Sweden. Fredrik.Lanner@ki.se.; Division of Obstetrics and Gynecology, Karolinska Universitetssjukhuset, Stockholm, Sweden. Fredrik.Lanner@ki.se.
Jazyk: angličtina
Zdroj: Nature communications [Nat Commun] 2020 Mar 30; Vol. 11 (1), pp. 1609. Date of Electronic Publication: 2020 Mar 30.
DOI: 10.1038/s41467-020-15326-5
Abstrakt: In vitro differentiation of human pluripotent stem cells into functional retinal pigment epithelial (RPE) cells provides a potentially unlimited source for cell based reparative therapy of age-related macular degeneration. Although the inherent pigmentation of the RPE cells have been useful to grossly evaluate differentiation efficiency and allowed manual isolation of pigmented structures, accurate quantification and automated isolation has been challenging. To address this issue, here we perform a comprehensive antibody screening and identify cell surface markers for RPE cells. We show that these markers can be used to isolate RPE cells during in vitro differentiation and to track, quantify and improve differentiation efficiency. Finally, these surface markers aided to develop a robust, direct and scalable monolayer differentiation protocol on human recombinant laminin-111 and -521 without the need for manual isolation.
Databáze: MEDLINE
Externí odkaz: