Detail of Fig8_R27-alpha-Cate



Project
Title
Time-lapse DIC images of R2/7 alpha-Cate cells culutured in non-adherent Lipidure-coated plates
Description
NA
Release, Updated
2017-10-03,
2018-11-15
License
CC BY
Kind
Image data based on Experiment
File Formats
Data size
32.5 MB

Organism
H. sapiens ( NCBITaxon:9606 )
Strain(s)
R2/7 alpha-Cate
Cell Line
-

Datatype
cell dynamics
Molecular Function (MF)
Biological Process (BP)
NA (spheroid formation, cell aggregate)
Cellular Component (CC)
cell ( GO:0005623 )
Biological Imaging Method
XYZ Scale
XY: 1.19 micrometer/pixel, Z: NA
T scale
30 minute for each time interval

Image Acquisition
Experiment type
TimeLapse
Microscope type
BrightfieldMicroscope
Acquisition mode
LaserScanningConfocalMicroscopy
Contrast method
Fluorescence
Microscope model
Olympus IX81
Detector model
Hamamatsu Photonics ORCA-ER
Objective model
Olympus UPlanFL N 10x/0.3 Ph1
Filter set
-

Summary of Methods
See details in Yonemura (2014) PLoS ONE, 9(11): e112922.
Related paper(s)

Shigenobu Yonemura (2014) Differential sensitivity of epithelial cells to extracellular matrix in polarity establishment., PloS one, Volume 9, Number 11, pp. e112922

Published in 2014 (Electronic publication in Nov. 13, 2014, midnight )

(Abstract) Establishment of apical-basal polarity is crucial for epithelial sheets that form a compartment in the body, which function to maintain the environment in the compartment. Effects of impaired polarization are easily observed in three-dimensional (3-D) culture systems rather than in two-dimensional (2-D) culture systems. Although the mechanisms for establishing the polarity are not completely understood, signals from the extracellular matrix (ECM) are considered to be essential for determining the basal side and eventually generating polarity in the epithelial cells. To elucidate the common features and differences in polarity establishment among various epithelial cells, we analyzed the formation of epithelial apical-basal polarity using three cell lines of different origin: MDCK II cells (dog renal tubules), EpH4 cells (mouse mammary gland), and R2/7 cells (human colon) expressing wild-type alpha-catenin (R2/7 alpha-Cate cells). These cells showed clear apical-basal polarity in 2-D cultures. In 3-D cultures, however, each cell line displayed different responses to the same ECM. In MDCK II cells, spheroids with a single lumen formed in both Matrigel and collagen gel. In R2/7 alpha-Cate cells, spheroids showed similar apical-basal polarity as that seen in MDCK II cells, but had multiple lumens. In EpH4 cells, the spheroids displayed an apical-basal polarity that was opposite to that seen in the other two cell types in both ECM gels, at least during the culture period. On the other hand, the three cell lines showed the same apical-basal polarity both in 2-D cultures and in 3-D cultures using the hanging drop method. The three lines also had similar cellular responses to ECM secreted by the cells themselves. Therefore, appropriate culture conditions should be carefully determined in advance when using various epithelial cells to analyze cell polarity or 3-D morphogenesis.
(MeSH Terms)

Contact
Shigenobu Yonemura , RIKEN , Center for Developmental Biology , Electron Microscope Laboratory
Contributors
Shigenobu Yonemura

OMERO Dataset
OMERO Project
Source