In-Vitro Development of Aggregates of Bovine Inner Cell Mass Cells or Bovine Mammary Cells and Putative Tetraploid Embryos Produced by Electrofusion.

Ito, Yasuko; Iwasaki, Setsuo

doi:10.1262/jrd.45.65

Cited by 9 publications

(9 citation statements)

References 13 publications

(19 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…They suggested that this may reflect the more committed nature of the ES cells to an ICM fate or simply their much smaller size. In our previous paper [11], when clumps of bovine mammary cells or bovine ICM stained with Hoechst were used for aggregation with bovine putative tetraploid embryos, all clumps contributed to the ICM. Recently Hadjantonakis et al [12] produced green fluorescent mice by germline transmission of green fluorescent ES cells.…”

Section: Discussionmentioning

confidence: 98%

GFP Expression in Mouse Fetuses Derived from Aggregation of Embryonic Stem Cells with Diploid or Tetraploid Embryos.

et al. 2000

Self Cite

View full text Add to dashboard Cite

Abstract:It is necessary to use a suitable marker gene to select embryos expressing transgene prior to transfer into recipient females especially in an efficient production of transgenic farm animals. Recently a green fluorescent protein (GFP) gene has been used as a marker gene for this purpose, because it is a self-fluorescent substance and does not require a substrate such as X-Gal in a lacZ gene. Using these characteristics of GFP, we transfected the GFP gene with the neomycinresistance gene under the control of the phosphoglycerase kinase promoter into mouse embryonic stem (ES) cells, and attempted to produce a transgenic mouse derived from aggregates of GFP-positive ES cells after the selection with G418 for 8 days with diploid or tetraploid embryos. There were some ES colonies expressing GFP at different levels after the selection with G418. These GFP-positive ES cells were used as a complete tight adhered clump for aggregation with diploid or tetraploid embryos and resulted in GFP expression in the inner cell mass (ICM) cells of aggregates, but no embryos expressing GFP in trophoblasts. At 9.5 days p.c., 6 out of 10 live fetuses (60%) for diploid⇔ES chimera, and all of 14 live fetuses (100%) for tetraploid⇔ ES chimera, in spite of head deformity, were transgenic fetuses. At 14.5 days p.c., among 7 normal embryos obtained from 21 embryos transferred to recipients, one fetus showed GFP expression in five organs (heart, kidney, liver, gonad and intestine), and 4 fetuses strongly expressed GFP in more than one organ as seen under the fluorescent dissecting microscope. These results indicate that enough transfected ES cells selected in a short time with G418 were able to be used for the selection of transgenic embryos prior to transfer to the recipient. Moreover, by using a complete tight adhered clump of ES cells for aggregation, the clump is able to completely contribute to a part of the ICM. The GFP gene may be useful and efficient as a marker especially for farm animals which have a long term pregnancy and a small litter size.

show abstract

Section: Discussionmentioning

confidence: 98%

GFP Expression in Mouse Fetuses Derived from Aggregation of Embryonic Stem Cells with Diploid or Tetraploid Embryos.

et al. 2000

Self Cite

View full text Add to dashboard Cite

show abstract

“…The tendency of ES cells to contribute to the embryonic lineages combined with tetraploid complementation of extraembryonic phenotypes may also be exploited to allow the production of embryos nearly 100% derived from ES cells . Although developed initially in the mouse, this technique has now been adopted in cattle (Iwasaki et al, 1999(Iwasaki et al, , 2000 and pigs (Prather et al, 1996).…”

Section: Tissue-specific Effects Of Tetraploidy: 4n:2n Mouse Chimerasmentioning

confidence: 99%

Tetraploid development in the mouse

Eakin

Behringer

2003

Developmental Dynamics

View full text Add to dashboard Cite

Spontaneous duplication of the mammalian genome occurs in approximately 1% of fertilizations. Although one or more whole genome duplications are believed to have influenced vertebrate evolution, polyploidy of contemporary mammals is generally incompatible with normal development and function of all but a few tissues. The production of tetraploid (4n) embryos has become a common experimental manipulation in the mouse. Although development of tetraploid mice has generally not been observed beyond midgestation, tetraploid:diploid (4n:2n) chimeras are widely used as a method for rescuing extraembryonic defects. The tolerance of tissues to polyploidy appears to be dependent on genetic background. Indeed, the recent discovery of a naturally tetraploid rodent species suggests that, in rare genetic backgrounds, mammalian genome duplications may be compatible with the development of viable and fertile adults. Thus, the range of developmental potentials of tetraploid embryos remains in large part unexplored. Here, we review the biological consequences and experimental utility of tetraploid mammals, in particular the mouse. Developmental Dynamics 228:751-766, 2003.

show abstract

“…On the other hand, bovine embryos fused at the two-cell stage developed to morulae or blastocysts with diploid/tetraploid mosaicism. Of a set of morulae developed after electrofusion 85.7% were tetraploid and 14.3% diploid (Iwasaki et al, 1999). In a recent study, even only 12.5% of bovine blastocysts produced by electrofusion of two-cell embryos were completely tetraploid and 41.7% displayed diploid/tetraploid mosaicism (Curnow et al, 2000).…”

mentioning

confidence: 96%

In vitro formation of tetraploid rat blastocysts after fusion of two‐cell embryos

Krivokharchenko

Galat

Ganten

et al. 2002

Molecular Reproduction Devel

View full text Add to dashboard Cite

Gene targeting technology is not available in the rat which is an animal model of major importance, e.g., in cardiovascular research. This is due to the fact that the rat embryonic stem cell (ESC)-like cells established by several groups do not form germ-line chimeras when injected into blastocysts. In the mouse, the aggregation of ESC with tetraploid embryos has allowed the generation of animals completely derived from these cells. However, aggregation of rat ESC-like cells with tetraploid rat embryos has not yet been attempted to evaluate their developmental capacity. Therefore, we established a method to produce tetraploid rat embryos by fusion at the two-cell stage. Chemical fusion by polyethylene glycol (PEG) was shown to be less efficient (56.3% fused embryos) than electrofusion (96.1% fused embryos). The rate of development of fused embryos to blastocysts was independent of the fusion method and similar to the rate of control embryos. However, this rate was lower when the embryos had been cultured from the zygote state before fusion (14-20%) compared to freshly isolated two-cell embryos (41-63%). Alike for the mouse, blastocysts derived from fused two-cell rat embryos contained about half the number of cells as control blastocysts and were homogeneously tetraploid with no evidence of mosaicism. This method may be useful for the establishment of gene-targeting technology in the rat.

show abstract

In-Vitro Development of Aggregates of Bovine Inner Cell Mass Cells or Bovine Mammary Cells and Putative Tetraploid Embryos Produced by Electrofusion.

Cited by 9 publications

References 13 publications

GFP Expression in Mouse Fetuses Derived from Aggregation of Embryonic Stem Cells with Diploid or Tetraploid Embryos.

GFP Expression in Mouse Fetuses Derived from Aggregation of Embryonic Stem Cells with Diploid or Tetraploid Embryos.

Tetraploid development in the mouse

In vitro formation of tetraploid rat blastocysts after fusion of two‐cell embryos

Contact Info

Product

Resources

About