photoreceptor development, retinal rod cell differentiation
Cellular Component (CC)
photoreceptor cilium
Biological Imaging Method
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XYZ Scale
XY:0.6251697 micrometer/pixel, Z: NA, XY:0.3125849 micrometer/pixel, Z: NA
T scale
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Image Acquisition
Experiment type
Immunofluorescence
Microscope type
ConfocalMicroscope
Acquisition mode
FluorescenceCorrelationSpectroscopy
Contrast method
Fluorescence
Microscope model
ZEISS LSM700
Detector model
-
Objective model
-
Filter set
-
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 )
(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.