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Biomedical subjects

Alan O Trounson

Publications and source records attributed to Alan O Trounson.

22 records · Page 2Linked to original sources

Human cloning 2001.

This review summaries human cloning from a clinical perspective. Natural human clones, that is, monozygotic twins, are increasing in the general community. Iatrogenic human clones have been produced for decades in infertile couples given fertility treatment such as ovulation induction. A clear distinction must be made between therapeutic cloning using embryonic stem cells and reproductive cloning attempts. Unlike the early clinical years of in vitro fertilization, with cloning there is no animal model that is safe and dependable. Until there is such a model, 'Dolly'-style human cloning is medically unacceptable.

Cloning, Organism↗

Glucose metabolism of human morula and blastocyst-stage embryos and its relationship to viability after transfer.

The pregnancy rate and implantation rate following blastocyst transfer in the human have been reported to be high; however, it has remained necessary to transfer 2-3 blastocysts to achieve these rates. Morphological criteria are currently used to select blastocysts for transfer and have some limited correlation with ongoing viability. Glucose metabolism of 189 human morula to blastocyst stage embryos was analysed using a non-invasive ultramicrofluorescence technique to determine if this could be used to predict viability. There was a linear trend to increased glucose uptake with progression from the morula to the hatching/hatched blastocyst stage of development, whereas glycolytic activity did not vary. There was no consistent difference in glucose uptake or glycolytic activity for embryos at the various morphological stages on day 5 compared to day 6 in vitro. Glucose uptake and glycolytic activity of the nine embryos positively identified as having implanted following transfer varied and were apparently not different from the values for embryos that failed to implant. In addition, viability was demonstrated to be compatible with high glycolytic activity, with four of nine implanted embryos having a glycolytic activity in the highest 15% of the population of embryos studied. Glucose uptake and glycolytic activity of male and female embryos did not appear to be different. Glucose metabolism cannot be used prospectively to select viable human morula or blastocyst stage embryos for transfer and it is also unlikely to be a useful tool to predict the sex of the embryo.

Journal Article↗

Human embryonic stem cells form embryoid bodies containing visceral endoderm-like derivatives.

OBJECTIVE: The objective of this study was to determine the potential of human embryonic stem (hES) cells to provide an in vitro model of human extraembryonic endoderm development. METHODS: The hES cell line HES-2 was propagated in Dulbecco's modification of Eagle's medium (DMEM) in the presence of 20% fetal calf serum (FCS) on a mouse embryonic fibroblast feeder layer. Clumps of approximately 50-100 cells were transferred to fresh DMEM and FCS and grown as embryoid bodies (EBs) in suspension culture. EBs were subjected to immunohistochemistry for endodermal, ectodermal and mesodermal specific markers and immunoreactivity analysed by confocal microscopy and on cryosections. RESULTS: HES-2 cells reproducibly formed spherical EBs after 2-3 days in suspension culture as clumps. EBs continued to expand and by 7 days had commenced cavitation in a highly reproducible and organised fashion. Culture periods longer than 10 days led to cystic structures displaying inconsistent morphological organisation. Immunolocalisation of anti-alpha-fetoprotein-, anti-neurofilament- and anti-CD31-specific antibodies at 7 days of culture provided evidence of regional differentiation of endodermal, ectodermal and mesodermal derivatives in cavitating EBs. Further, of 10 cavitating EBs analysed at 7 days of culture, all displayed immunolocalisation of anti-pan-keratin-, anti-CK8- and anti-alpha-fetoprotein-specific antibodies to a peripheral cellular layer, suggestive of yolk sac visceral endoderm (VE) formation. CONCLUSION: Cavitation, the presence of regionalised cell lineage-specific immunoreactivity and the development of a VE-like peripheral cell layer demonstrate that the HES-2 hES cell line can be induced to undergo EB formation and provide scope to study early human primitive endoderm and yolk sac VE development.

Animals↗