SAMN19844094
External Id
Zhejiang University
INSDC center name
2022-12-13T00:50:13.750Z
INSDC first public
2022-12-13T00:50:13.750Z
INSDC last update
SRS9276500
INSDC secondary accession
live
INSDC status
Model organism or animal
NCBI submission model
Model.organism.animal.1.0
NCBI submission package
SRS9276500
SRA accession
8-week-old
age
Shanghai SLAC Laboratory
biomaterial_provider
crypt cell
cell_type
Core facilities, Zhejiang university school of medicine
collected_by
Epithelial cells were isolated from the small intestine. In brief, isolated small intestines were opened lengthwise, washed with cold PBS. After removal of villi by scraping with a cold glass slide, samples were chopped into pieces of around 2 mm and further washed with cold PBS. The tissue was then incubated in 2.5 mM EDTA with PBS at 4 degree Celsius for 30 min on a rocking platform at 20 rpm. The supernatant was then removed and the tissue pieces were resuspended in DMEM/F12 (Corning) with 0.1% BSA, shaken vigorously and then filtered through a 70 um strainer (Corning) whilst on ice. Crypts were collected by centrifugation at 4 degree Celsius and then digested with TrypLE Express enzyme 1X (Gibco) and 0.5 mg/ml DNase (Worthington) at 37 degree Celsius for 5 min before filtration through a 40 um strainer (Corning). Single-cell suspensions were prepared as above and resuspended in FACS buffer containing HBSS with 25mM HEPES (Gibco) and 1% fetal bovine serum (Gibco). Antibody staining was performed at 4 degree Celsius for 30 min. 7-AAD (BD Biosciences, 51-68981E) was added to the final suspensions before isolation of required single cells by FACS (Beckman moflo Astrios EQ). Cells (7-AAD-CD45-EpCAM+CD44+) sorted by FACS were then subjected to single-cell sequencing using a 10X Genomics platform.
description
Mus musculus
http://purl.obolibrary.org/obo/NCBITaxon_10090
organism
pooled male and female
sex
C57BL/6
strain
the small intestine
tissue
Crypt cells from the adult mouse small intestine
title
Cells (7-AAD-CD45-EpCAM+CD44+) sorted by FACS
treatment