Detail of Fig1g_DD11
Project
Title
Bright field microscopy images of pigmented cells from Nrl-GFP mouse iPSC (induced pluripotent stem cells) at differentiation day (DD) 11
Description
NA
Release, Updated
2017-10-03,
2018-11-15
2018-11-15
License
CC BY
Kind
Image data
based on Experiment
File Formats
Data size
9.2 MB
Gene symbols
Nrl
Protein tags
GFP
Datatype
cell/tissue structure
Molecular Function (MF)
Biological Process (BP)
-
Cellular Component (CC)
cell
(
GO:0005623
)
Biological Imaging Method
XYZ Scale
XY: 3.24 micrometer/pixel, Z: NA
T scale
-
Image Acquisition
Experiment type
Other
Microscope type
Other
Acquisition mode
Other
Contrast method
Fluorescence
Microscope model
a motorized inverted system microscope (IX71, Olympus)
Detector model
charge-coupled device camera (DP72, Olympus), and digital imaging software (DP2-BSW, Olympus)
Objective model
Olympus UPlanFL 4 × 0.13 phL
Filter set
-
Summary of Methods
See details in Iwasaki et al. (2016) PLoS ONE, 11(7): e0158282.
Related paper(s)
Yuko Iwasaki, Sunao Sugita, Michiko Mandai, Shigenobu Yonemura, Akishi Onishi, Shin-Ichiro Ito, Manabu Mochizuki, Kyoko Ohno-Matsui, Masayo Takahashi (2016) Differentiation/Purification Protocol for Retinal Pigment Epithelium from Mouse Induced Pluripotent Stem Cells as a Research Tool., PloS one, Volume 11, Number 7, pp. e0158282
Published in 2016 (Electronic publication in July 6, 2016, midnight )
- doi: 10.1371/journal.pone.0158282
-
pmid: 27385038
(Abstract) PURPOSE: To establish a novel protocol for differentiation of retinal pigment epithelium (RPE) with high purity from mouse induced pluripotent stem cells (iPSC). METHODS: Retinal progenitor cells were differentiated from mouse iPSC, and RPE differentiation was then enhanced by activation of the Wnt signaling pathway, inhibition of the fibroblast growth factor signaling pathway, and inhibition of the Rho-associated, coiled-coil containing protein kinase signaling pathway. Expanded pigmented cells were purified by plate adhesion after Accutase(R) treatment. Enriched cells were cultured until they developed a cobblestone appearance with cuboidal shape. The characteristics of iPS-RPE were confirmed by gene expression, immunocytochemistry, and electron microscopy. Functions and immunologic features of the iPS-RPE were also evaluated. RESULTS: We obtained iPS-RPE at high purity (approximately 98%). The iPS-RPE showed apical-basal polarity and cellular structure characteristic of RPE. Expression levels of several RPE markers were lower than those of freshly isolated mouse RPE but comparable to those of primary cultured RPE. The iPS-RPE could form tight junctions, phagocytose photoreceptor outer segments, express immune antigens, and suppress lymphocyte proliferation. CONCLUSION: We successfully developed a differentiation/purification protocol to obtain mouse iPS-RPE. The mouse iPS-RPE can serve as an attractive tool for functional and morphological studies of RPE.(MeSH Terms)
- Animals
- Cell Communication
- Cell Culture Techniques[major]
- Cell Differentiation[major]
- Cell Proliferation
- Cells, Cultured
- Collagenases/chemistry
- Embryonic Stem Cells/cytology
- Induced Pluripotent Stem Cells/cytology[major]
- Lymphocytes/cytology
- Mice
- Peptide Hydrolases/chemistry
- Phagocytosis
- Retinal Pigment Epithelium/metabolism[major]
- Rod Cell Outer Segment/metabolism
- Signal Transduction
- rho-Associated Kinases/metabolism
Contact
Masayo Takahashi
, RIKEN
, Center for Developmental Biology
, Laboratory for Retinal Regeneration
Contributors
Yuko Iwasaki, Sunao Sugita, Michiko Mandai, Shigenobu Yonemura, Akishi Onishi, Shin-ichiro Ito, Manabu Mochizuki Kyoko Ohno-Matsui, Masayo Takahashi
OMERO Dataset
OMERO Project