Sample 2
Protocols: After incubation, cells were rinsed in PBS twice before being lysed directly in Buffer RLT Plus from the RNeasy plus micro kit (Qiagen, Victoria, Australia). Cells were cultured in a humidified incubator at 37ºC, with 5% carbon dioxide. Growth medium provided contains 1 g/L D-glucose (Gibco, Victoria, Australia) Dulbecco's Modified Eagle Medium (DMEM), L-Glutamine and 110 mg/L Sodium pyruvate, supplemented with 10% foetal bovine serum (Bovogen, Victoria, Australia) and 1% Penicillin-Streptomycin (Gibco, Victoria, Australia)  Cells were cultured on either collagen biomaterial or tissue culture plastic at 1,000,000 cells per sample collection over 48 hours in growth medium.  Total RNA was extracted from lysed cells using the RNeasy Plus Micro kit (Qiagen, Victoria, Australia). Quality of RNA was determined on the 2100 BioAnalyzer (Agilent Technologies, Victoria, Australia). All samples of at least an RNA Integrity Number (RIN) of ≥ 8.0 and an A260/280 ratio of 1.8 – 2.0 were accepted. Library preparation was performed on 1 µg total RNA per sample with the Illumina TruSeq® Stranded mRNA kit according to manufacturer's protocol (Illumina, San Diego, California).
isolate
not applicable
organism
Homo sapiens
developmental stage
adult
disease
normal
growth condition
collagen biomaterial
cell_type
adipose tissue derived mesenchymal stem cell
replicate
technical replicate
ENA-FIRST-PUBLIC
2021-12-23
ENA-LAST-UPDATE
2021-12-23