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An alternative method of deriving embryonic stem cell-like clones by aggregation of diploid cells with tetraploid embryos
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- Authors
- Issue Date
- 2006
- Publisher
- Elsevier
- Citation
- Fertility and Sterility, vol.85 no.1, pp. 1103-1110
- Keywords
- Mouse embryo ; tetraploidy ; aggregation ; embryonic stem (ES) cell ; characterization
- Abstract
- Objective: To assess whether embryonic stem (ES) cells could be derived from the aggregation of diploid cells
with tetraploid embryos.
Design: Randomized, prospective study.
Setting: University embryology and gamete biotechnology laboratory.
Animal(s): F1 (C57BL6/DBA2) mice.
Intervention(s): Four- to eight-cell F1 tetraploid embryos were aggregated with 10 to 15 donor E14 ES cells.
Main Outcome Measure(s): Embryogenesis and ES cell establishment.
Result(s): No difference (78% to 89%) in blastocyst formation was detected between the aggregated tetraploid and the
control diploid embryos. In a total of 27 transfers, pregnancy was detected in three tetraploid (23.1%) and five diploid
(35.7%) cases, and three live births developed from the aggregated tetraploid embryos. The tetraploid blastocysts
without aggregation were plated, but no ES cell-like colony was formed. Six of eight aggregated blastocysts derived
well-proliferated colonies, which were positive for anti–stage-specific embryonic antigen (SSEA)-1 antibody, Oct-4,
and alkaline phosphatase. The microsatellite assay confirmed the homogenous makeup among the donor E14 cells and
live-birth and ES-like cells derived from the E14-aggregated, tetraploid embryo.
Conclusion(s): The aggregation of pluripotent diploid cells with tetraploid embryos yielded live births and ES-like
cells that were homogenous to the donor diploid cells.
- ISSN
- 0015-0282
- Language
- English
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