Detail of DD20_Sop_RXRg_EtOH
Project
Title
Immunofluorescence image of photoreceptor-specific gene S-opsin and RXRy expressing in mouse iPS cells derived 3D retinal maintained containing EtOH at differentiated day20
Description
NA
Release, Updated
2019-11-20
License
CC BY
Kind
Image data
based on Experiment
File Formats
Data size
3.2 MB
Gene symbols
S-opsin, RXRy
Datatype
retina differentiation
Molecular Function (MF)
Biological Process (BP)
retinal rod cell differentiation
(
GO:0060221
)
photoreceptor development
(
GO:0008594
)
Cellular Component (CC)
photoreceptor cilium
(
GO:0032391
)
Biological Imaging Method
XYZ Scale
XY:0.3125849 micrometer/pixel, Z: NA
T scale
-
Image Acquisition
Experiment type
Immunofluorescence
Microscope type
ConfocalMicroscope
Acquisition mode
FluorescenceCorrelationSpectroscopy
Contrast method
Fluorescence
Microscope model
ZEISS LSM700
Detector model
-
Objective model
-
Filter set
-
Summary of Methods
See details in Ueda et al. (2018) Biochem Biophys Res Commun, 495(4): 2595-2601.
Related paper(s)
Kaori Ueda, Akishi Onishi, Shin-Ichiro Ito, Makoto Nakamura, Masayo Takahashi (2018) Generation of three-dimensional retinal organoids expressing rhodopsin and S- and M-cone opsins from mouse stem cells., Biochemical and biophysical research communications, Volume 495, Number 4, pp. 2595-2601
Published in 2018 Jan 22 (Electronic publication in Dec. 20, 2017, midnight )
- doi: 10.1016/j.bbrc.2017.12.092
-
pmid: 29274337
(Abstract) PURPOSE: Three-dimensional retinal organoids can be differentiated from embryonic stem cells/induced pluripotent stem cells (ES/iPS cells) under defined medium conditions. We modified the serum-free floating culture of embryoid body-like aggregates with quick reaggregation (SFEBq) culture procedure to obtain retinal organoids expressing more rod photoreceptors and S- and M-cone opsins. METHODS: Retinal organoids differentiated from mouse Nrl-eGFP iPS cells were cultured in various mediums during photoreceptor development. To promote rod photoreceptor development, organoids were maintained in media containing 9-cis retinoic acids (9cRA). To obtain retinal organoids with M-opsin expression, we cultured in medium with 1% fetal bovine serum (FBS) supplemented with T3, BMP4, and DAPT. Section immunohistochemistry was performed to visualize the expression of photoreceptor markers. RESULTS: In three-dimensional (3D) retinas exposed to 9cRA, rhodopsin was expressed earlier and S-cone opsins were suppressed. We could maintain 3D retinas up to DD 35 in culture media with 1% FBS. The 3D retinas expressed rhodopsin, S- and M-opsins, but most cone photoreceptors expressed either S- or M-opsins. CONCLUSION: By modifying culture conditions in the SFEBq protocol, we obtained rod-dominated 3D retinas and S- and M-opsin expressing 3D retinas.(MeSH Terms)
- Animals
- Bioartificial Organs[major]
- Cell Differentiation/physiology
- Cells, Cultured
- Cone Opsins/metabolism[major]
- Mice
- Organogenesis/physiology
- Printing, Three-Dimensional
- Retina/cytology[major]
- Retina/growth & development[major]
- Retinal Cone Photoreceptor Cells/metabolism
- Rhodopsin/metabolism[major]
- Stem Cells/cytology[major]
- Stem Cells/physiology
- Tissue Engineering/methods[major]
Contact
Akishi Onishi
, RIKEN
, Center for Biosystems Dynamics Research
, Laboratory for Retinal Regeneration
Contributors
Kaori Ueda, Akishi Onishi, Shin-ichiro Ito, Makoto Nakamura, Masayo Takahashi
OMERO Dataset
OMERO Project