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 uterus. Here we show that cell lines can be derived from the epiblast, a tissue of the post-implantation embryo that generates the embryo proper. These cells, which we refer to as EpiSCs (post-implantation epiblast-derived stem 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.
Recent reports have suggested that mammalian stem cells residing in one tissue may have the capacity to produce differentiated cell types for other tissues and organs 1-9. Here we define a mechanism by which progenitor cells of the central nervous system can give rise to non-neural derivatives. Cells taken from mouse brain were co-cultured with pluripotent embryonic stem cells. Following selection for a transgenic marker carried only by the brain cells, undifferentiated stem cells are recovered in which the brain cell genome has undergone epigenetic reprogramming. However, these cells also carry a transgenic marker and chromosomes derived from the embryonic stem cells. Therefore the altered phenotype does not arise by direct conversion of brain to embryonic stem cell but rather through spontaneous generation of hybrid cells. The tetraploid hybrids exhibit full pluripotent character, including multilineage contribution to chimaeras. We propose that transdetermination consequent to cell fusion 10 could underlie many observations otherwise attributed to an intrinsic plasticity of tissue stem cells 9.
A suspension is made in isotonic (2.2%) sodium citrate solution from the contents of the tubules from a whole testis or a testicular biopsy specimen. The germinal cells are sedimented by centrifuging, leaving most of the sperm in the supernatant fluid, which is discarded. The cells are resuspended in hypo-tonic (1%) sodium citrate solution and left to stand at room temperature for 12 minutes, after which they are sedimented again and fixed as a concentrated suspension in a mixture of 3 parts absolute ethyl alcohol to 1 part glacial acetic acid plus a trace of chloroform. Two quick changes into fresh fixative follow. Air-dried preparations are made from the final fixed suspension and stained in lactic-acetic-orcein. The method is suitable for stages of male meiosis in which the chromosomes are condensed. Its principle advantage is the separation of the clumps of spermatogonia and spermatocytes into individual cells which are randomly dispersed over the preparations. Compared with squash techniques, the air-drying method gives improved spreading of the chromosomes and less cell breakage.
a b s t r a c tHuman activities have profoundly changed the land on which we live. In particular, land use and land management change affect the hydrology that determines flood hazard, water resources (for human and environmental needs) and the transport and dilution of pollutants. It is increasingly recognised that the management of land and water are inextricably linked (e.g. Defra, 2004). "Historical context, state of the science and current management issues" section of this paper addresses the science underlying those linkages, for both rural and urban areas. In "Historical context, state of the science and current management issues" section we discuss future drivers for change and their management implications. Detailed analyses are available for flood risk, from the Foresight Future Flooding project (Evans et al., 2004a,b) and other recent studies, and so we use flooding as an exemplar, with a more limited treatment of water resource and water quality aspects. Finally in "Science needs and developments" section we discuss science needs and likely progress. This paper does not address the important topic of water demand except for some reference to the Environment Agency's Water Resources Strategy for England and Wales (Environment Agency, 2009).
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