Effect of the Polyvinylpyrrolidone Concentration of Cryoprotectant on Mouse Embryo Development and Production of Pups: 7.5% of PVP is Beneficial for In Vitro and In Vivo Development of Frozen-Thawed Mouse Embryos

Kim, Chi-Gu; Yong, Hae In; Lee, Gene; Cho, Jaejin

doi:10.1262/jrd.19185

Cited by 16 publications

(8 citation statements)

References 34 publications

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“…1A). Briefly, follicles were released by puncturing the ovaries in M2 medium [46]. They were then incubated for 15 minutes at 37°C in 0.25% trypsin (Gibco-BRL, Carlsbad, CA), 0.2% collagenase IV (Gibco-BRL) plus 0.02% DNase-I (Sigma) solution, and finally repeatedly pipetted with a drawn pipette.…”

Section: Methodsmentioning

confidence: 99%

Growth of Mouse Oocytes to Maturity from Premeiotic Germ Cells In Vitro

et al. 2012

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In the present study, we established an in vitro culture system suitable for generating fertilizable oocytes from premeiotic mouse female germ cells. These results were achieved after first establishing an in vitro culture system allowing immature oocytes from 12–14 day- old mice to reach meiotic maturation through culture onto preantral granulosa cell (PAGC) monolayers in the presence of Activin A (ActA). To generate mature oocytes from premeiotic germ cells, pieces of ovaries from 12.5 days post coitum (dpc) embryos were cultured in medium supplemented with ActA for 28 days and the oocytes formed within the explants were isolated and cocultured onto PAGC monolayers in the presence of ActA for 6–7 days. The oocytes were then subjected to a final meiotic maturation assay to evaluate their capability to undergo germinal vesicle break down (GVBD) and reach the metaphase II (MII) stage. We found that during the first 28 days of culture, a significant number of oocytes within the ovarian explants reached nearly full growth and formed preantral follicle-like structures with the surrounding somatic cells. GSH level and Cx37 expression in the oocytes within the explants were indicative of proper developmental conditions. Moreover, the imprinting of Igf2r and Peg3 genes in these oocytes was correctly established. Further culture onto PAGCs in the presence of ActA allowed about 16% of the oocytes to undergo GVBD, among which 17% reached the MII stage during the final 16–18 hr maturation culture. These MII oocytes showed normal spindle and chromosome assembly and a correct ERK1/2 activity. About 35% of the in vitro matured oocytes were fertilized and 53.44% of them were able to reach the 2-cell stage. Finally, around 7% of the 2-cell embryos developed to the morula/blastocyst stage.

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Section: Methodsmentioning

confidence: 99%

Growth of Mouse Oocytes to Maturity from Premeiotic Germ Cells In Vitro

et al. 2012

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“…On the other hand, Checura and Seidel (2007) reported the successful vitrification of bovine oocytes with 20 and 6% PVP [44]. Mouse embryo was successfully vitrified with supplementation of EG and sucrose as a cryoprotectant solution with 7.5% PVP [45].…”

Section: Discussionmentioning

confidence: 99%

Effect of Polyvinylpyrrolidone on Vitrification of Buffalo (Bubalus bubalis) Oocytes

Bari¹,

Islam²,

Alam³

et al. 2020

J. Buffalo Sci.

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Vitrification, a method of rapid cooling, is an alternate cryopreservation method of oocytes and embryos. The present study was aimed to examine the effect of polyvinylpyrrolidone (PVP) on vitrification of buffalo oocytes. Cumulus oocyte complexes (COCs) with fully grown oocytes (120-130 µm in diameter) were aspirated from slaughtered buffalo ovaries for vitrification. COCs were treated with equilibration solution at room temperature for 5 min and then transferred to a vitrification solution for 1 min. Then the COCs were submerged into liquid nitrogen (-196C) for a while using cryotops. The COCs were thawed, diluted, and washed in a washing solution for 5 min, respectively. Vitrified oocytes were incubated for in vitro maturation (IVM) at 38.5C under an atmosphere of 5% CO2 in the air for 24 hrs. Cumulus cells surrounding the oocytes were removed mechanically, oocytes were fixed in acetic acid and ethanol, and stained with aceto-orcein to examine the meiotic stages of oocytes. The numbers of morphologically normal oocytes after vitrification were higher in 5% PVP than 0 and 10% PVP groups. A proportion of oocytes treated with 5% PVP reached the metaphase II (MII) stage while none of the oocytes from 0% and 10% PVP groupsdeveloped beyond anaphase I and metaphase I (MI) stages, respectively. These results show that PVP can be used as a cryoprotectant for the vitrification of buffalo oocytes.

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“…Sucrose (0.07 M final concentration) and Y-27632, a selective inhibitor of the Rho-associated protein kinase p160 ROK (10 mM final conc., Calbiochem, San Diego CA, USA), were also evaluated for their added ability to safeguard SSCs through the freezing process (Gauthaman et al, 2010). In addition, the cryoprotectant activity of polyvinylpyrrolidone (1% PVP; Sigma, cat# P-0930) was assessed as a replacement for FBS (Kim et al, 2008). To investigate the effect that different freezing conditions have on SSC viability, thawed testis cells were transplanted into recipient testes as described above.…”

Section: Cryopreservation Studiesmentioning

confidence: 99%

Fertile Offspring Derived from Mouse Spermatogonial Stem Cells Cryopreserved for More Than 14 Years.

Wu¹,

Goodyear²,

Abramowitz³

et al. 2012

Biology of Reproduction

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background: Approximately 80% of childhood cancers can now be cured but a side effect of treatment results in about one-third of the surviving boys being infertile or severely subfertile when they reach reproductive age. Currently, more than 1 in 5000 men of reproductive age who are childhood cancer survivors suffer from this serious quality of life problem. It is possible to obtain a testicular biopsy before treatment to preserve the spermatogonial stem cells (SSCs) of the male by cryopreservation, but the results of long-term storage of SSCs on their subsequent functional ability to generate normal offspring has not been examined in any mammalian species. Moreover, it will be necessary to increase the number of these cryopreserved SSCs to remove any contaminating malignant cells and assure regeneration of spermatogenesis. methods and results: In this report, we demonstrate that long-term cryopreservation (.14 years) of testis cells from mouse, rat, rabbit and baboon safeguards SSC viability, and that these cells can colonize the seminiferous tubules of recipient testes. Moreover, mouse and rat SSCs can be cultured and re-establish complete spermatogenesis, and fertile mouse progeny without apparent genetic or epigenetic errors were generated by the sperm produced.conclusions: These findings provide a platform for fertility preservation in prepubertal boys undergoing gonadotoxic treatments.

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Effect of the Polyvinylpyrrolidone Concentration of Cryoprotectant on Mouse Embryo Development and Production of Pups: 7.5% of PVP is Beneficial for In Vitro and In Vivo Development of Frozen-Thawed Mouse Embryos

Cited by 16 publications

References 34 publications

Growth of Mouse Oocytes to Maturity from Premeiotic Germ Cells In Vitro

Growth of Mouse Oocytes to Maturity from Premeiotic Germ Cells In Vitro

Effect of Polyvinylpyrrolidone on Vitrification of Buffalo (Bubalus bubalis) Oocytes

Fertile Offspring Derived from Mouse Spermatogonial Stem Cells Cryopreserved for More Than 14 Years.

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