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

Plaza Reyes, Alvaro ; Petrus-Reurer, Sandra ; Padrell Sánchez, Sara ; Kumar, Pankaj ; Douagi, Iyadh ; Bartuma, Hammurabi LU ; Aronsson, Monica ; Westman, Sofie ; Lardner, Emma and André, Helder , et al. (2020) In Nature Communications 11(1).
Abstract

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.... (More)

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.

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publishing date
type
Contribution to journal
publication status
published
subject
keywords
Animals, Biomarkers/metabolism, CD56 Antigen, Cell Differentiation/physiology, Embryonic Stem Cells, Epithelial Cells/metabolism, Humans, Laminin/genetics, Macular Degeneration/metabolism, Neurons/metabolism, Rabbits, Retinal Pigment Epithelium/metabolism, Retinal Pigments/metabolism
in
Nature Communications
volume
11
issue
1
pages
15 pages
publisher
Nature Publishing Group
external identifiers
  • pmid:32231223
  • scopus:85082731796
ISSN
2041-1723
DOI
10.1038/s41467-020-15326-5
language
English
LU publication?
no
id
4603111f-191e-46f1-8ef2-ca0799066d36
date added to LUP
2021-08-09 14:28:05
date last changed
2024-04-06 06:24:39
@article{4603111f-191e-46f1-8ef2-ca0799066d36,
  abstract     = {{<p>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.</p>}},
  author       = {{Plaza Reyes, Alvaro and Petrus-Reurer, Sandra and Padrell Sánchez, Sara and Kumar, Pankaj and Douagi, Iyadh and Bartuma, Hammurabi and Aronsson, Monica and Westman, Sofie and Lardner, Emma and André, Helder and Falk, Anna and Nandrot, Emeline F and Kvanta, Anders and Lanner, Fredrik}},
  issn         = {{2041-1723}},
  keywords     = {{Animals; Biomarkers/metabolism; CD56 Antigen; Cell Differentiation/physiology; Embryonic Stem Cells; Epithelial Cells/metabolism; Humans; Laminin/genetics; Macular Degeneration/metabolism; Neurons/metabolism; Rabbits; Retinal Pigment Epithelium/metabolism; Retinal Pigments/metabolism}},
  language     = {{eng}},
  month        = {{03}},
  number       = {{1}},
  publisher    = {{Nature Publishing Group}},
  series       = {{Nature Communications}},
  title        = {{Identification of cell surface markers and establishment of monolayer differentiation to retinal pigment epithelial cells}},
  url          = {{https://lup.lub.lu.se/search/files/101032682/Identification_of_cell_surface_markers.pdf}},
  doi          = {{10.1038/s41467-020-15326-5}},
  volume       = {{11}},
  year         = {{2020}},
}