Similarly to what was done using the leukemia cell lines, to study the effects of BMSCs on CD34 cells, gene ex pression profiles from CD34 cell mono cultures and co cultures with BMSCs have been analyzed by microarrays. We found that 2075 genes were differentially expressed in CD34 cell co cultures compared with mono cultures. Among essentially the most up regulated genes had been SOCS3, REN and CXCL6, all having a fold transform 5. Ingenu ity pathway analysis with the differentially expressed genes re vealed that probably the most represented canonical pathways had been cAMP mediated signaling, VDR RXR activation and car or truck diac B adrenergic signaling. CCL2 and IL 8 are elevated in supernatants from BMSCs co cultured with leukemia cells Gene expression evaluation revealed that the majority of the genes up regulated in BMSCs co cultured with leukemia cells were involved in IL 17 signaling.
To assess the variables created by co cultured cells, we screened over here the superna tants from co culture and mono culture samples at 48 h for cytokine production by R D Human Cytokine panel A. We chose this panel because amongst the 36 cytokines in the panel were CXCL1, sICAM 1, IL 1B, IL eight, CCL2 and Serpin E1 all of which were identified to be up regulated at the gene level in co cultured BMSCs. Additionally, with panel A we had been in a position to measure the relative levels of IFN?, IL 6 and IL 23 that are IL 17 signaling connected cytokines. Nevertheless, the majority of the 36 cytokines in the panel have been undetectable in our samples plus the levels of cyto kines CXCL1, ICAM 1, IL 23, IL 6, MIF and Serpin E1 were not substantially changed between co culture and mono culture situations.
However, the levels of CCL2 and IL 8 had been higher in supernatants from BMSCs co cultured with leukemia cells, however the final results were variable among BMSCs from various subjects. The levels of IFN? selleck inhibitor and CD40L were higher in co culture compared with mono culture supernatants, however the differ ence was not statistically important. The evaluation of cyto kines inside the supernatant of cultured BMSCs and leukemia cells was performed in three series of experiments with BMSCs from three healthful donors, and we found distinct responses among the unique BMSC donors. We located enhanced levels of IFN? and CD40L only inside the supernatants from BMSC003 co cultured with TF 1 and TF1. The levels of IL 8 had been elevated in the supernatants from BMSC003, BMSC006 and, to a lesser extent, in BMSC002. The amount of CCL2 was measurable only in supernatants from BMSC003, BMSC002 and BMSC006 co cultured with TF 1 and K562 leukemia cells and inside the supernatant from BMSC006 co cultured with TF 1. To confirm the increased levels of CCL2 and IL eight inside the supernatants from BMSCs co cultured with leukemia cells at 48 h, we measured the levels of the two cytokines applying ELISA assays.