Detail of Fig1F



Project
Title
Time-lapse fluorescence images of of the three-dimensional-retinas from Fstl4::Venus mouse embryonic stem cell (ESC) from differentiation day (DD) 10 to 20
Description
NA
Release, Updated
2018-11-14
License
CC BY
Kind
Image data based on Experiment
File Formats
Data size
15.0 MB

Organism
M. musculus ( NCBITaxon:10090 )
Strain(s)
ES
Cell Line
-
Gene symbols
Fstl4
Protein tags
Venus

Datatype
tissue structure
Molecular Function (MF)
Biological Process (BP)
-
Cellular Component (CC)
-
Biological Imaging Method
XYZ Scale
-
T scale
-

Image Acquisition
Experiment type
TimeLapse
Microscope type
ConfocalMicroscope
Acquisition mode
LaserScanningConfocalMicroscopy
Contrast method
Fluorescence
Microscope model
Olympus IX71
Detector model
-
Objective model
-
Filter set
-

Summary of Methods
See details in Maekawa et al. (2015) Curr Eye Res, 41(4): 558-568.
Related paper(s)

Yuki Maekawa, Akishi Onishi, Keizo Matsushita, Naoshi Koide, Michiko Mandai, Kiyoshi Suzuma, Takashi Kitaoka, Atsushi Kuwahara, Chikafumi Ozone, Tokushige Nakano, Mototsugu Eiraku, Masayo Takahashi (2016) Optimized Culture System to Induce Neurite Outgrowth From Retinal Ganglion Cells in Three-Dimensional Retinal Aggregates Differentiated From Mouse and Human Embryonic Stem Cells., Current eye research, Volume 41, Number 4, pp. 558-68

Published in 2016 Apr (Electronic publication in April 16, 2015, midnight )

(Abstract) PURPOSE: To establish a practical research tool for studying the pathogenesis of retinal ganglion cell (RGC) diseases, we optimized culture procedures to induce neurite outgrowth from three-dimensional self-organizing optic vesicles (3D-retinas) differentiated in vitro from mouse and human embryonic stem cells (ESCs). MATERIALS AND METHODS: The developing 3D-retinas isolated at various time points were placed on Matrigel-coated plates and cultured in media on the basis of the 3D-retinal culture or the retinal organotypic culture protocol. The number, length, and morphology of the neurites in each culture condition were compared. RESULTS: First, we confirmed that Venus-positive cells were double-labeled with a RGC marker, Brn3a, in the 3D-retina differentiated from Fstl4::Venus mouse ESCs, indicating specific RGC-subtype differentiation. Second, Venus-positive neurites grown from these RGC subsets were positive for beta-III tubulin and SMI312 by immunohistochemistry. Enhanced neurite outgrowth was observed in the B27-supplemented Neurobasal-A medium on Matrigel-coated plates from the optic vesicles isolated after 14 days of differentiation from mouse ESCs. For the differentiated RGCs from human ESCs, we obtained neurite extension of >4 mm by modifying Matrigel coating and the culture medium from the mouse RGC culture. CONCLUSION: We successfully optimized the culture conditions to enhance lengthy and high-frequency neurite outgrowth in mouse and human models. The procedure would be useful for not only developmental studies of RGCs, including maintenance and projection, but also clinical, pathological, and pharmacological studies of human RGC diseases.
(MeSH Terms)

Contact
Akishi Onishi , RIKEN , Center for Developmental Biology , Laboratory for Retinal Regeneration
Contributors
Yuki Maekawa, Akishi Onishi, Keizo Matsushita, Naoshi Koide, Michiko Mandai, Kiyoshi Suzuma, Takashi Kitaoka, Atsushi Kuwahara, Chikafumi Ozone, Tokushige Nakano, Mototsugu Eiraku, Masayo Takahashi

OMERO Dataset
OMERO Project
Source