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Development of reconstituted embryos derived from transgenic embryonic stem cell nuclei

Content Provider	Springer Nature Link
Author	Hosaka, Koji Sato, Kahei
Copyright Year	2002
Abstract	This study was carried out to transform embryonic stem (ES) cells and to produce the reconstituted embryos derived from transgenic ES cell nuclei. Then, in vitro/vivo developmental potency of transgenic ES cells were compared to that of control ES cells (non-transgenic ES cells) in the reconstituted embryos.Unfertilized B6D2F1 ooplasm at metaphase II (M II) and two kinds of ES cell lines, 129SV and transgenic (tg) 129SV transformed by EGFP gene, were used as nuclear recipients and nuclear donors, respectively. The M II chromosome-spindle complex was aspirated into the pipette with a minimal volume of ooplasm. After enucleation, the ES cell nuclei was injected into the enucleated ooplasm directly. Then, reconstituted embryos were activated in SrCl$_{2}$, and they were cultured in HTF medium.There was no difference of developmental rate between reconstituted embryos derived from the control (non-transgenic) and the tg ES cells. From this result, we indicated that transgenic ES cells might not change the property of peculiarity of the ES cell by gene transfer. The expression of GFP gene in the embryos was observed by fluorescence microscope at the 4-cell and more stage. As comparison with development of the embryos derived from the control and tg ES cells, the difference of the development could not be confirmed between the two cell groups. When the reconstituted embryos derived from the control and tg ES cells were transferred into oviduct or uterus of pseudopregnant females, fetuses were observed 13.5 days post coitum. However, in all fetuses, developmental arrest and regression were seen 19.5 days post coitum.
Starting Page	224
Ending Page	229
Page Count	6
File Format	PDF
ISSN	09147470
Journal	Human Cell
Volume Number	15
Issue Number	4
e-ISSN	17490774
Language	English
Publisher	Springer-Verlag
Publisher Date	2012-08-03
Publisher Place	Tokyo
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	clone mouse nuclear transfer ES cell transgenic mouse embryo Cell Biology Embryology Oncology Stem Cells Reproductive Medicine Cell Culture
Content Type	Text
Resource Type	Article
Subject	Cell Biology Medicine Cancer Research

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