Gene expression of E13 fetal liver of common lymphoid progenitors Flt3+ a4b7+ compared to a4b7-HSAint, a4b7-HSAhi or a4b7-HSAlow

We used whole genome transcriptome as gene discovery to further understand the rules of lineage restriction in the lymphoid compartment Mouse embryos at days 13 gestation were isolated and the fetal livers were dissected. Single cell suspension was prepared and treated with biotinilated antibodies to CD3, CD4, CD8, TER119, Gr1, CD25, CD19, NK1.1 and CD11c. Cell suspensions were then incubated with streptavidin Miltenyi Macs beads. Lineage negative cells were recovered in Macs collumns. Cell suspensions were then incubated with antibodies against Flt3, c-kit, Sca-1, a4b7, HSA and Streptavidin bound to Pacific Blue. Lin-, c-kit+, Flt3+, Sca-1lo, a4b7+ or a4b7-HSAlow,or a4b7- HSAint or a4b7- HSAhi cells were then isolated by cell sorting. 3 biological replicates of each sample were isolated with 5x103 cells. These subsets were shown in single cell diferentiation assays to be have differentpotentials because a4b7+ cells lost B cell potential, while HSA low partially lost B cell potential, HSAhi partially lost T cell potential and HSA int are unbiased in their differentiation potential.