Stem cell factor (SCF) effect on mouse bone marrow derived dendritic cells (BMdDCs) under hypoxia

BMdDCs were cultured under hypoxic conditions for 2 days and then for additional 6 hours either in the presence or absence of SCF at 100 ng/ml. Differentially expressed genes were evaluated by Affymetrix microarray (Affymetrix Mouse Gene 2.1 ST Array). Four identical experiments were performed (experiments were named as follows: BC; D2; E; I). Each time BMdDCs were generated under normoxia from mouse bone marrow cells by in vitro culture with GM-CSF for 7 days and subsequent CD11c+ cell-purification by CD11c-microbeads (4 biological replicates of BMdDCs). Then highly purified CD11c+ (>98% pure) BMdDCs were cultured under hypoxia (2% O2, ~14 mmHg) in the following conditions: with GM-CSF at 20 ng/ml for 2 days, and finally either in the presence or absence of SCF at 100 ng/ml for further 6 hours (medium and SCF conditions, respectively).