Project description:Hypoblast stem cells were derived from rat embryonic stem cells (rESCs) by culturing with LIF, EGF, PDGF, and 2% fetal bovine serum. mRNA expression profiles of the cells during derivation at different time points were determined with Affymetrix microarrays. Global transcriptome of rESC-derived hypoblast stem cells (cHypoSCs) was similar to that of rat embryo-derived hypoblast stem cells and rat multipotent adult progenitor cells. In addition, the transcriptome of these cells were compared with the previously published datasets including transcriptome of mouse embryonic stem cells, extraembryonic endoderm cells, single cells of mouse embryos (E3.25, E3.5, and E4.5). The comparison revealed that cHypoSCs have the transcriptome similar to that of extraembryonic endoderm cells and E3.5 primitive endoderm cells.
Project description:We report the mRNA profiles of single cells with high-throughput sequencing (RNA-seq) of hypoblast stem cells derived from rat embryonic stem cells. The single-cell data was combined with the previously published dataset: transcriptomes of mouse embryo cells (inner cell mass or trophectoderm). The clustering revealed that rat embryonic stem cell-derived hypoblast stem cells were similar to the mouse inner cell mass cells.
Project description:Rat Extraembryonic Endoderm Precursor (XEN-P) cells resemble the nascent hypoblast of the blastocyst. Here we show that rat Multipotent Adult Progenitor Cell (rMAPC) lines derived from bone marrow (BM) exhibit salient nascent hypoblast characteristics. We could not identify cells with hypoblast features in rat BM, but they appeared after prolonged in vitro culture under rMAPC conditions. However, rMAPC culture conditions supported the very rapid isolation of new lines with nascent hypoblast features from rat blastocysts that were more homogenous than XEN-P cells isolated on feeders. The gene expression profile, growth factor requirement, Oct4 promoter methylation, and in vitro/vivo differentiation potential of cells isolated using rMAPC culture conditions from BM or blastocysts or XEN-P cells were highly similar. We conclude that the three cell populations have characteristics of hypoblast stem cells (HypoSC) and that rMAPC culture conditions support the isolation of XEN-P/HypoSC from the blastocyst and induction of HypoSC from BM. Comparison between bone marrow and blastocyst-derived Hypoblast Stem Cells (hypoSC) Three cell lines derived from bone marrow, three cell lines derived from blastocyst and one mesenchymal used, as control (for all of them 2 replicates)
Project description:Rat Extraembryonic Endoderm Precursor (XEN-P) cells resemble the nascent hypoblast of the blastocyst. Here we show that rat Multipotent Adult Progenitor Cell (rMAPC) lines derived from bone marrow (BM) exhibit salient nascent hypoblast characteristics. We could not identify cells with hypoblast features in rat BM, but they appeared after prolonged in vitro culture under rMAPC conditions. However, rMAPC culture conditions supported the very rapid isolation of new lines with nascent hypoblast features from rat blastocysts that were more homogenous than XEN-P cells isolated on feeders. The gene expression profile, growth factor requirement, Oct4 promoter methylation, and in vitro/vivo differentiation potential of cells isolated using rMAPC culture conditions from BM or blastocysts or XEN-P cells were highly similar. We conclude that the three cell populations have characteristics of hypoblast stem cells (HypoSC) and that rMAPC culture conditions support the isolation of XEN-P/HypoSC from the blastocyst and induction of HypoSC from BM.