CD54+BP1+YFP- bone marrow stromal cells from IL7 reporter mice

Hematopoiesis in adult mammals involves cognate interactions between developing hematopoietic cells and bone marrow stromal cell niches. SCF and CXCL12 play key roles in the maintenance of HSC, while early B cell differentiation requires CXCL12 and IL7. In this study, we characterized mouse BM stromal cells expressing IL7 by performing a transcriptomic analysis. We found that SCF, CXCL12 and IL7 were co-expressed by a unique peri-sinusoidal stromal cell subset. CD45-Lin-CD54+BP1+ and CD45-Lin-YFP+CD54+BP1+ cells were isolated by flow cytometry from the bone marrow of 6 to 12 wo C57Bl/6J and IL7-Cre/Rosa-eYFP mice respectively (1800 to 10000 cells). Total bone marrow cells from C57Bl/6 mice were used as a control. Total RNA was amplified using the NuGEN Ovation Pico WTA System V2 Kit. cDNA hybridization was performed on Affymetrix Mouse GE ST 1.0 microarrays. CD45-Lin-CD54+BP1+YFP- cells were isolated by flow cytometry from the bone marrow of 6 to 12 wo IL7-Cre/Rosa-eYFP mice (2300 cells). Total RNA was amplified using the NuGEN Ovation Pico WTA System V2 Kit. cDNA hybridization was performed on Affymetrix Mouse GE ST 1.0 microarrays.