RNaseq of primary human OA chondrocytes alones or in transwell cocultures with primary human Adipose-derived Mesenchymal Stromal Cells, in presence of 10nM rapamycin or DMSO as vehicle control
The objective was to study the combined effects of rapamycin and primary human Adipose-derived Mesenchymal Stromal Cells (AD-MSC) on primary human OA chondrocytes (versus their effects separetely), in an in-vitro model that reproduce an intra-articular injection.   For this purpose, P1 OA chondrocytes were seeded in 6-well plates (5x10^5 cells/well)  in their proliferative medium  for 24h. In parallel, P1 AD-MSC were seeded in 0.4 µm inserts (7.2x10^4) in their proliferative medium for 24h. The next days, all media were removed, cells were washed twice with PBS and media were replaced by miminal chondrogenic medium. Finally, OA chondrocytes were cocultured with AD-MSC (versus alone) in presence of 10nM rapamycin or DMSO (at 1:10000 dilution) as vehicle control for 3 days. At the end, media and inserts were removed and RNA of OA chondrocytes were extracted using RNeasy kit from QIAGEN with an on-column DNase I digestion as manufacturer's instructions.   Because of poor quality RNA, one sample (25_DMSO_AD10) and his control (25_RA10_AD10) have been excluded from this analysis. Otherwise, all RNA integrity numbers were above 9 before libraries construction.