Project description:We used whole genome transcriptome as gene discovery to further understand the differential behaviour of the first (embryonic) and second waves of thymic settling progenitors in the murine embryo.
Project description:Here we compared the expression of an engineered kidney tissue, created by recombining an in vitro budded Wolffian duct with fresh E13 metanephric mesenchyme, with that of three in vivo rat embryonic kidney timepoints (E13, E18, and week 4) Keywords: time course
Project description:We used whole genome transcriptome as gene discovery to further understand the differential behaviour of the first (embryonic) and second waves of thymic settling progenitors in the murine embryo. Mouse embryos at days 13 and 18 of gestation were isolated and the thymic lobes 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 thymocytes were recovered in Macs columns. Cell suspensions were then incubated with antibodies against CD135, CD117, CD44, CD24 and Streptavidin bound to PECy7. Lin-, CD117+, CD44+, CD24low, CD135+ cells were then isolated by cell sorting. 3 biological replicates of each sample were isolated with 104-3x104 cells
Project description:Here we compared the expression of an engineered kidney tissue, created by recombining an in vitro budded Wolffian duct with fresh E13 metanephric mesenchyme, with that of three in vivo rat embryonic kidney timepoints (E13, E18, and week 4) Experiment Overall Design: The global gene expression of recombined tissue (non-branched in vitro-formed UBs and MM) was analyzed and compared to the gene expression of early, late, and post developmental kidneys in order to determine whether normal developmental pathways were being followed. Genes with at least a 3-fold difference in expression between any of the 4 conditions (E13, E18, Wk4, recombined tissue) were analyzed and grouped into one of ten expression patterns. The expression of each group of genes was analyzed in the recombined tissue to compare the recombined tissue gene expression levels to the three in vivo time points.
