Detail of DD30_Rod_CRX_EtOH
Project
Title
Immunofluorescence image of photoreceptor-specific gene crx and Rhodopsin expressing in mouse iPS cells derived 3D retinal maintained containing EtOH at differentiated day30
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
crx, Rhodopsin
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