RNAseq of primary human Adipose-derived Mesenchymal Stromal Cells stimulated for 2 days with 20ng/mL IFN-gamma (versus unstimulated control) in presence of 10 nM rapamycin or DMSO (at 1:10000 dilution) as vehicle control
In order to study the impact of rapamycin on AD-MSC under inflammatory or resting conditions, primary human Adipose-derived Mesenchymal Stromal Cells (AD-MSC) at passage 1 were plated at 1.5x10^4 in MEM-alpha, 5% Human Platelet Lysate, 1% Penicillin-Streptomycin and allowed to stand for 24h. The next day, AD-MSC were stimulated for 2 days (or not) with 20ng/mL IFN-gamma in presence of 10nM rapamycin (from 10.000X stock in DMSO) or DMSO (at 1:10000 dilution) as vehicle control. After 2 days, media were removed and RNA were extracted using RNeasy kit from QIAGEN with an on-column DNAse I digestion as manufacturer's instructions. RNA intergrity number were estimated on Agilent Bioanalyzer and were all above 9 before sequencing. Libraries and RNA sequencing were performed by the Beijing Genomics Institute (BGI)