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