Project description:Direct pairwise comparison of the differential expression patterns between embryos injected with Standard olignucleotide morpholinoes and those injected with an anti-klf4 morpholino. Keywords: Direct pairwise comparison
Project description:3 different ES cell lines were compared in order to determine whether there are significant expression profile differences between ES cell lines, or whether the constraints of maintaining pluripotency in culture force a similar expression profile on cell lines derived from disparate sources. Our results indicate that the latter is more likely. We identified 21 genes that were significantly differentially regulated, either on comparison with the pooled control, or on direct comparison of individual ES cell line data from different slides. Using semi-quantitative RT-PCR on 3 separate isolates from each cell line, we have confirmed 4 of these genes as consistently differentially regulated, Hprt, and 3 others. We would conclude therefore that different ES cell lines at the same passage number in identical culture conditions show very similar expression profiles. Keywords: cell type comparison
Project description:Homozygous disruption of Bteb2/Klf5, a homolog of Drosophila gap gene Krüppel, led to increased expression of various differentiation marker genes, such as Fgf5, Cdx2, and Brachyury in mouse ES cells without compromising their ability to differentiate into all three germ layers. Upon removal of LIF, Klf5-deficient ES cells showed faster differentiation kinetics than wild-type ES cells. In contrast, overexpression of Klf5 in ES cells suppressed the transcription of differentiation marker genes, and maintained pluripotency in the absence of LIF. In order to search downstream genes of Klf5, we surveyed genes implicated in ES cell proliferation by microarray analysis Keywords: cell type comparison
Project description:Pairwise comparison of total kidney RNA extracted from Nas1 knockout mice versus total kidney RNA extracted from wildtype mice Keywords: Direct pairwise comparison
Project description:Direct comparison of FACS sorted EGFP positive brain macrophages vs pooled Total Brain extract Keywords: Direct pairwise comparison
Project description:Direct comparison of FACS sorted EGFP positive renal macrophages vs pooled Total Kidney extract Keywords: Direct pairwise comparison
Project description:Embryonic stem cells (ES cells) are pluripotent stem cells that can contribute to all lineages of the embryo proper and were first isolated in 1981 in this laboratory (Evans and Kaufmann, 1981). They have contributed immeasurably to biology, both in terms of studying pluripotent stem cell maintenance in culture and in vitro differentiation, and because they can be easily genetically manipulated in culture, allowing for the production of mice with designed specific mutations (targeted mice), via homologous recombination. The recent isolation of human ES cell lines is promising in terms of stem cell therapy development. Mouse ES cell lines are isolated from the inner cell masses of mouse blastocysts between days 3.5 and 5.0 post coitum, or from the inner cell masses of delayed blastocysts. However, it has been suggested that some aspects of their biology may be more similar to embryonic ectoderm from early post implantation embryos, than to pre-implantation blastocysts. It has also been suggested that human embryonic stem cells are more similar to mouse stem cells derived from early embryonic ectoderm, than to mouse ES cells. Until recently, however, it has been difficult to answer these questions definitively, as the quantity of tissue available from early embryos is extremely limiting. However, we have used two rounds of RNA amplification on early embryonic samples and individual colonies picked from ES cell culture to compare the transcriptomic profiles of mouse ES cells with the embryonic tissues from with these cells are commonly derived, and with slightly later embryonic stages. We have found that ES cells are notably most similar in profile to embryonic ectoderm at day 5.5 (EE5.5) . ES cells cannot be derived from this post implantation embryonic tissue, so this result is in some ways surprising. However, ES cells show key differences from EE5.5, including the expression of markers of pluripotency such as Oct4 and Nanog (see gene lists acompanying this record), which explain how their ability to contribute to all tissues of the embryo proper is retained. Keywords: cell type comparison
Project description:The application of human embryonic stem (ES) cells in medicine and biology has an inherent reliance on understanding the starting cell population. Human ES cells differ from mouse ES cells and the specific embryonic origin of both cell types is unclear. Previous work suggested that mouse ES cells could only be obtained from the embryo before implantation in the uterus1–5. Here we show that cell lines can be derived from the epiblast, a tissue of the postimplantation embryo that generates the embryo proper. These cells, which we refer to as EpiSCs (post-implantation epiblastderived ES cells), express transcription factors known to regulate pluripotency, maintain their genomic integrity, and robustly differentiate into the major somatic cell types as well as primordial germ cells. The EpiSC lines are distinct from mouse ES cells in their epigenetic state and the signals controlling their differentiation. Furthermore, EpiSC and human ES cells share patterns of gene expression and signalling responses that normally function in the epiblast. These results show that epiblast cells can be maintained as stable cell lines and interrogated to understand how pluripotent cells generate distinct fates during early development. *Note: EpiSCs were previously referred to as post-ES cells Keywords: cell type comparison