2024-07-26
ENA first public
ERC000011
ENA-CHECKLIST
SAMEA114498717
External Id
UOM
INSDC center name
2023-10-16T13:13:23Z
INSDC last update
public
INSDC status
ERS16480370
SRA accession
E-MTAB-13435:Sample 11
Submitter Id
64
year
age
ArrayExpress
broker name
not collected
collection date
human
common name
Protocols: All experiments and procedures carried out in this study were performed with relevant approval from the National Research Ethics Service and The University of Manchester. Adipose tissue was obtained from donors undergoing hip replacement surgery, with each donor providing full written informed consent. The adipose derived stem cells (ASC) extraction process was undertaken as previously described [PMID: 32489577]. ll experiments and procedures carried out in this study were performed with relevant approval from the National Research Ethics Service and The University of Manchester. Adipose tissue was obtained from donors undergoing hip replacement surgery, with each donor providing full written informed consent. The adipose derived stem cells (ASC) extraction process was undertaken as previously described [PMID: 32489577]. Donor ASCs used for experimentation were expanded in culture medium containing: 110mg/L sodium pyruvate, 1000 mg/L glucose and further supplemented with final concentrations of 10% (v/v) FBS, 2mM GlutaMAX (Life Technologies), 50μg/mL ascorbate, 100U/mL penicillin, 100μg/mL streptomycin and 0.25μg/mL amphotericin (subsequently termed standard culture medium). ASCs were grown in ambient incubator conditions (37°C, 5% CO2, 20% O2) until sufficient cell numbers were obtained. All donors were used between passages 3-5 and around 14 days in culture. ASCs were seeded in flasks or multiwell plates at a density of 7.5x103 cells/cm2 (70-80% confluence) and allowed to adhere for 24hrs. Cells were washed twice in PBS and serum starved in FBS-free standard culture medium (SFM) for a further 24hrs. On the day of GDF6 stimulation, medium was aspirated, cells washed twice in PBS and stimulated with 100ng/mL of GDF6 (Peprotech) in SFM as previously described [PMID: 32489577]. Samples were taken at 4 timepoints (0h, 2h, 6h, and 12h). Total RNA was extracted using TRIzol according to the manufacturers protocol, then resuspended in 20µL TE buffer. Libraries were generated using the TruSeq® Stranded mRNA assay (Illumina, Inc.) according to the manufacturer’s protocol
description
adult
developmental stage
not available
disease
not collected
geographic location (country and/or sea)
Donor3
individual
not applicable
isolate
Homo sapiens
organism
Homo sapiens
organism
adipose tissue
organism part
Homo sapiens
scientific_name
female
sex
GDF6 stimulation
stimulus
Sample 11
title