O-156. Survival and in-vitro implantation of interphase-stage and mitotic-stage 1-cell mouse embryos after vitrification

AbstractIn multicellular organisms, oocytes and sperm undergo fusion during fertilization and the resulting zygote gives rise to a new individual. The ability of zygotes to produce a fully formed individual from a single cell when placed in a supportive environment is known as totipotency. Given that totipotent cells are the source of all multicellular organisms, a better understanding of totipotency may have a wide-ranging impact on biology. The precise delineation of totipotent cells in mammals has remained elusive, however, although zygotes and single blastomeres of embryos at the two-cell stage have been thought to be the only totipotent cells in mice. We now show that a single blastomere of two- or four-cell mouse embryos can give rise to a fertile adult when placed in a uterus, even though blastomere isolation disturbs the transcriptome of derived embryos. Single blastomeres isolated from embryos at the eight-cell or morula stages and cultured in vitro manifested pronounced defects in the formation of epiblast and primitive endoderm by the inner cell mass and in the development of blastocysts, respectively. Our results thus indicate that totipotency of mouse zygotes extends to single blastomeres of embryos at the four-cell stage.

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Development of early somitic mouse embryos in static culture in vitro

Journal of Experimental Zoology ◽

10.1002/jez.1402200210 ◽

1982 ◽

Vol 220 (2) ◽

pp. 219-225 ◽

Cited By ~ 5

Author(s):

Yu-Jui Wan ◽

Tsung-Chieh Wu ◽

Ivan Damjanov

Keyword(s):

Mouse Embryos ◽

Culture In Vitro ◽

Static Culture

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Peri-implantation mouse embryos: An in vitro assay for assessing serum-associated embryotoxicity in women with reproductive disorders

Reproductive Toxicology ◽

10.1016/s0890-6238(97)00144-5 ◽

1998 ◽

Vol 12 (2) ◽

pp. 155-159 ◽

Cited By ~ 5

Author(s):

Amos Fein ◽

Howard Carp ◽

Arkady Torchinsky ◽

Mara Koifman ◽

Ronit Yacobovich ◽

...

Keyword(s):

In Vitro Assay ◽

Mouse Embryos ◽

Reproductive Disorders ◽

Vitro Assay

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Effect of co-culturing of half-destroyed and intact 4-cell mouse embryos in varying ratios on subsequent in vitro development

Theriogenology ◽

10.1016/j.theriogenology.2010.12.031 ◽

2011 ◽

Vol 75 (9) ◽

pp. 1682-1687 ◽

Cited By ~ 1

Author(s):

U. Sanmee ◽

W. Piromlertamorn ◽

T. Vutyavanich

Keyword(s):

Mouse Embryos ◽

In Vitro Development ◽

Vitro Development

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Quality assessment of nine paracetamol 500 mg tablet brands marketed in Saudi Arabia

BMC Research Notes ◽

10.1186/s13104-021-05672-y ◽

2021 ◽

Vol 14 (1) ◽

Author(s):

Reem AlSwayeh ◽

Syed N. Alvi ◽

Muhammad M. Hammami

Keyword(s):

Saudi Arabia ◽

Active Substance ◽

Immediate Release ◽

Water Medium ◽

Breaking Force ◽

Weight Variation ◽

Percent Difference ◽

Stage 1

Abstract Objective To evaluate in-vitro quality of paracetamol 500 mg tablet brands marketed in Saudi Arabia. Results Two reference (R1 and R2) and seven generic (G1-G7) brands were commercially available. Four brands were single-drug, containing paracetamol only (R1, G1-G3) and five contained additional active ingredients (R2, G4-G7). All brands were immediate-release. Weight variation (n = 20, range as percent difference from mean), active substance content (n = 20, mean (SD) as percent difference from label), breaking force (n = 10, mean (SD)), and friability (n = 20, as percent weight loss) ranged from 97 to 102%, 96.1% (2.9%) to 99.8% (1.1%), 9.9 (0.4) to 21.0 (0.9) kg, and 0.017% to 0.809%, respectively. Disintegration (water medium) time (n = 6, minute: second) ranged from 02:35–03:09 to 12:49–13:10. Dissolution (phosphate buffer, pH 5.8) profile showed a mean release at 30 min of 87% to 97% of label content, with seven brands passing stage-1 (≥ 85% for each of 6 test units) and two passing stage-2 (mean of 12 test units ≥ 85%) criteria. Despite statistically significant differences between R1 and R2 and some of their corresponding generic brands in active substance content, breaking force, and amount dissolved at 30 min, all nine brands met the pre-specified quality standards.

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Mouse chick chimera: a new model to study the in ovo developmental potentialities of mammalian somites

Development ◽

10.1242/dev.121.6.1705 ◽

1995 ◽

Vol 121 (6) ◽

pp. 1705-1718 ◽

Cited By ~ 7

Author(s):

J. Fontaine-Perus ◽

V. Jarno ◽

C. Fournier le Ray ◽

Z. Li ◽

D. Paulin

Keyword(s):

In Vitro Cultures ◽

Mouse Embryos ◽

Limb Muscle ◽

Mammalian Development ◽

In Ovo ◽

Gene Coding ◽

Myogenic Stem Cells ◽

Mouse Cells ◽

Beta Galactosidase

Chimeras were prepared by transplanting somites from 9-day post-coitum mouse embryos or somitic dermomyotomes from 10-day post-coitum mouse embryos into 2-day-old chick embryos at different axial levels. Mouse somitic cells then differentiated in ovo in dermis, cartilage and skeletal muscle as they normally do in the course of development and were able to migrate into chick host limb. To trace the behavior of somitic myogenic stem cells more closely, somites arising from mice bearing a transgene of the desmin gene linked to a reporter gene coding for Escherichia coli beta-galactosidase (lacZ) were grafted in ovo. Interestingly, the transgene was rapidly expressed in myotomal muscles derived from implants. In the limb muscle mass, positive cells were found several days after implantation. Activation of desmin nls lacZ also occurred in in vitro cultures of somite-derived cells. Our experimental method facilitates investigation of the mechanisms of mammalian development, allowing the normal fate of implanted mouse cells to be studied and providing suitable conditions for identification of descendants of genetically modified cells.

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Diploid and haploid mouse parthenogenetic development following in vitro activation and embryo transfer

Development ◽

10.1242/dev.31.3.635 ◽

1974 ◽

Vol 31 (3) ◽

pp. 635-642

Author(s):

M. H. Kaufman ◽

R. L. Gardner

Keyword(s):

Embryo Transfer ◽

Mouse Embryos ◽

Parthenogenetic Development ◽

In Vitro Activation ◽

Haploid Embryos ◽

Parthenogenetic Mouse Embryos

Parthenogenetic mouse embryos were selected following in vitro activation, and transferred to the oviducts of pseudopregnant recipients. Decidua was evoked by 50–56% of diploid parthenogenones compared to 35·1% of haploid embryos with a single pronucleus, 37·5% of immediate cleavage eggs and 77% of fertilized eggs (controls). On day 4, 58·7% of diploid parthenogenones were morphologically normal morulae or blastocysts; over 90% of these ‘normal’ embryos evoked decidua when retransferred to recipients compared to 8·9% of abnormal embryos flushed from the ‘transfer’ sides, suggesting that only ‘normal’ embryos could evoke decidua. Potentially diploid parthenogenones remained diploid on chromosomal examination on day 4.

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Modification of cell division by phorbol ester in preimplantation mouse embryos

Development ◽

10.1242/dev.101.2.403 ◽

1987 ◽

Vol 101 (2) ◽

pp. 403-408

Author(s):

E.T. Mystkowska ◽

W. Sawicki

Keyword(s):

Cell Division ◽

Video Recording ◽

Mouse Embryos ◽

Time Intervals ◽

Cell Divisions ◽

Stage Of Development ◽

Preimplantation Mouse Embryos ◽

Preimplantation Mouse ◽

Single Blastomeres

2-cell mouse embryos were treated in vitro with a 2 h pulse of phorbol myristate acetate (PMA) at 32nd, 38th and 50th h after hCG, then chased in culture for up to 46 h. Embryos were fixed at various time intervals of chasing, then stained and inspected. Some embryos were carefully inspected with a video recording system, every 1.44s and the cell divisions (cytokinesis) as well as formation of large, single blastomeres, each from two smaller ones, were recorded. PMA pulse let to the suppression of cell divisions. The rate of the suppression was time dependent: with a delay of 0–1, 12 and 18 h between the PMA pulse and time of scheduled cell division about 99, 87 and 44% of 2-cell embryos remained at this stage of development, for at least 10 h, respectively, and 90, 58 and 12% of their blastomeres revealed binuclearity. Since we found that PMA-mediated formation of binuclearity was not the effect of cell fusions, it was assumed that the inhibition of cytokinesis preceded by karyokinesis was responsible for binuclearity. PMA effect on cell divisions was reversible. PMA-treated embryos revealed formation of large, single blastomeres, each from two smaller ones. If cell division appeared after PMA pulse, in about 52% of 3- to 6-cell embryos, the large blastomere formation was recorded in the course of the subsequent 38 h. Large blastomere formation was concluded to be the result of either cell fusion or reversion of incompleted cytokinesis brought about by PMA.

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