Sample 1
Homo sapiens
Protocols: Biopsy was obtained from the patient under informed consent in DMEM F12 containing 10% FBS and 2% P/S and processed within 2 hours. The tissue was minced into small pieces and washed extensively with PBS before being dissociated with collagenase IV for 2-3 hours and additional 15' with TripLE at 37C. Digestion was stopped by medium dilution and the pellet was resuspended in 70% Matrigel. Organoids were cultured in endometrial cancer organoids culture medium. organoids were cultured embedded in 70% Matrigel in 48-wells low attachment culture plates in endometrial cancer organoid culture medium which was refreshed every 2 days. Organoids were enzymatically and mechanically passaged with TripLE every 2 weeks. In order to be passaged, organoids were removed from Matrigel with ice-cold DMEM F12 and centrifuge at 230g for 5'. The pellet was resuspended in 500ul of TripLE and incubated for 10' at 37C. Incubation was stopped by dilution and organoids were mechanically tritured before being centrifuged at 230g for 5'. Finally, the pellet was resuspended in 70% Matrigel and cells were seeded. Matrigel was allowed to solidify and 250ul of pre-warmed medium was added thereafter. Organoids were split at 1/4 to 1/10 ratio. Organoids with low passage numbers were used for this experiments unless otherwise stated. Organoids were recovered from Matrigel with ice-cold DMEM F12 and centrifuged at 230g for 5'. The pellet was resuspended in 200ul of PBS and genomic DNA was isolated using the Purelink Genomic Mini Kit (Invitrogen) according to the manufacturer's instructions. DNA concentration was measured with a nanodrop and DNA was stored at -80C before being processed for exome sequencing. Whole genome DNA libraries were created using the Illumina TruSeq DNA sample preparation kit V2 according to the manufacturer's instructions.
ENA-FIRST-PUBLIC
2019-06-04T17:04:07Z
ENA-LAST-UPDATE
2019-02-14T17:39:24Z
External Id
SAMEA5340601
INSDC center name
VIB Center for Cancer Biology (CCB) Laboratory for Translational Genetics Department of Human Genetics KULeuven
INSDC first public
2019-06-04T17:04:07Z
INSDC last update
2019-02-14T17:39:24Z
INSDC status
public
Submitter Id
E-MTAB-7687:Sample 1
age
not available
broker name
ArrayExpress
common name
human
developmental stage
adult
disease
Endometrial cancer
disease staging
I
growth condition
organoid culture
individual
1
organism part
endometrium
sample name
E-MTAB-7687:Sample 1
scientific_name
Homo sapiens
sex
female
time
primary
tumor grading
IA