Human leukocyte expression profiles (in vitro)

CD34+ progenitor cells were cultured for 0 or 21 days in StemPro medium + supplement + IL-3 at 5 ng/ml Basophils, eosinophils and plasmacytoid dendritic cells were purified to near homogeneity and lysed for miRNA and mRNA. Basophils were purified by negative selection (StemSep Abs, miltenyi columns), eosinophils by StemSep Abs and pDC by positive selection after removing monocytes. The method starting with a 2-step Percoll gradient that put eosinophils in the pellet, basophils in the middle layer and pDC in the upper layer. Cells were isolated from purified to high homegeneity (99%) and lysed for RNA.