Promotion of haematopoietic activity in embryonic stem cells by the aorta-gonad-mesonephros microenvironment.

We investigated whether the in vitro differentiation of ES cells into haematopoietic progenitors could be enhanced by exposure to the aorta-gonadal-mesonephros (AGM) microenvironment that is involved in the generation of haematopoietic stem cells (HSC) during embryonic development. We established a co-culture system that combines the requirements for primary organ culture and differentiating ES cells and showed that exposure of differentiating ES cells to the primary AGM region results in a significant increase in the number of ES-derived haematopoietic progenitors. Co-culture of ES cells on the AM20-1B4 stromal cell line derived from the AGM region also increases haematopoietic activity.

Severe global DNA hypomethylation blocks differentiation and induces histone hyperacetylation in embryonic stem cells.

It has been reported that DNA methyltransferase 1-deficient (Dnmt1-/-) embryonic stem (ES) cells are hypomethylated (20% CpG methylation) and die through apoptosis when induced to differentiate. Here, we show that Dnmt[3a-/-,3b-/-] ES cells with just 0.6% of their CpG dinucleotides behave differently: the majority of cells within the culture are partially or completely blocked in their ability to initiate differentiation, remaining viable while retaining the stem cell characteristics of alkaline phosphatase and Oct4 expression.

Interpretation with hindsight.

In his 1965 paper 'DNA content of tumours: cytophotometric measurements', Sandritter was for the first time able to relate tumour DNA content to the pathology and progression of a small number of tumours. In subsequent publications, these observations were extended to the progression of a much more comprehensive range of tumours. The interpretation of Sandritter's paper below follows the increasing sophistication of methodologies for tumour DNA content through the existing publications and evaluates the conclusions and hypotheses Sandritter proposed in the light of the contemporary account.

Expression of a novel homeobox gene Ehox in trophoblast stem cells and pharyngeal pouch endoderm.

Ehox is an X-linked paired like homeobox gene identified from a differentiating embryonic stem (ES) cell cDNA library and is expressed at low levels in the preimplantation blastocyst and in ES cells in vitro. In embryos at 6.5 days post coitum (dpc), Ehox expression was restricted to the extraembryonic ectoderm which correlates with high-level expression in cultures of trophoblast stem cells.

Quantitative developmental anatomy of definitive haematopoietic stem cells/long-term repopulating units (HSC/RUs): role of the aorta-gonad-mesonephros (AGM) region and the yolk sac in colonisation of the mouse embryonic liver.

In the developing mouse embryo the first definitive (transplantable-into-the-adult) haematopoietic stem cells/long-term repopulating units (HSC/RUs) emerge in the AGM region and umbilical vessels on 10-11 days post coitum (d.p.c.

Cloning and characterization of Ehox, a novel homeobox gene essential for embryonic stem cell differentiation.

We report here the identification and characterization of a novel paired-like homeobox-containing gene (Ehox). This gene, identified in embryonic stem (ES) cells, is differentially expressed during in vitro ES cell differentiation. We have assessed Ehox function using the ES cell in vitro differentiation system.

DNA ligase I null mouse cells show normal DNA repair activity but altered DNA replication and reduced genome stability.

DNA ligase I is the key ligase for DNA replication in mammalian cells and has also been reported to be involved in a number of recombination and repair processes. Our previous finding that Lig1 knockout mouse embryos developed normally to mid-term before succumbing to a specific haematopoietic defect was difficult to reconcile with a report that DNA ligase I is essential for the viability of cultured mammalian cells. To address this issue, we generated a second Lig1 targeted allele and found that the phenotypes of our two Lig1 mutant mouse lines are identical.