Efficient collection and cryopreservation of embryos in F344 strain inbred rats

Taketsuru, Hiroaki; Kaneko, Takehito

doi:10.1016/j.cryobiol.2013.07.004

Cited by 26 publications

(14 citation statements)

References 24 publications

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“…Then, pronuclear-stage embryos were collected from the superovulated females previously mated with males. They were cultured in a modified Krebs-Ringer bicarbonate solution48 before and after the microinjections49. Using a micromanipulator (Narishige, Tokyo, Japan), 100 ng μl −1 Cas9 mRNA and 50 ng μl −1 gRNA were microinjected into the male pronuclei of the embryos.…”

Section: Methodsmentioning

confidence: 99%

Allele-specific genome editing and correction of disease-associated phenotypes in rats using the CRISPR–Cas platform

et al. 2014

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The bacterial CRISPR/Cas system has proven to be an efficient gene-targeting tool in various organisms. Here we employ CRISPR/Cas for accurate and efficient genome editing in rats. The synthetic chimeric guide RNAs (gRNAs) discriminate a single-nucleotide polymorphism (SNP) difference in rat embryonic fibroblasts, allowing allele-specific genome editing of the dominant phenotype in (F344 × DA)F1 hybrid embryos. Interestingly, the targeted allele, initially assessed by the allele-specific gRNA, is repaired by an interallelic gene conversion between homologous chromosomes. Using single-stranded oligodeoxynucleotides, we recover three recessive phenotypes: the albino phenotype by SNP exchange; the non-agouti phenotype by integration of a 19-bp DNA fragment; and the hooded phenotype by eliminating a 7,098-bp insertional DNA fragment, evolutionary-derived from an endogenous retrovirus. Successful in vivo application of the CRISPR/Cas system confirms its importance as a genetic engineering tool for creating animal models of human diseases and its potential use in gene therapy.

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Section: Methodsmentioning

confidence: 99%

Allele-specific genome editing and correction of disease-associated phenotypes in rats using the CRISPR–Cas platform

et al. 2014

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“…It has been reported that superovulation in Wistar and F344 rats can be induced by injecting PMSG and hCG [1, 15]. On the contrary, it is empirically known that the number of BN oocytes that ovulated by induction of superovulation is extremely low.…”

Section: Discussionmentioning

confidence: 99%

“…Female Wistar, F344, and BN rats aged 8 to 12 weeks were induced to superovulate by intraperitoneal injection of PMSG (ASKA Pharmaceutical, Tokyo, Japan) at doses of 150, 150, and 300 IU/kg, respectively, at 1600–1700 h, followed by intraperitoneal injection of hCG (ASKA Pharmaceutical) at doses of 75, 75, and 300 IU/kg, respectively, 48 h later [15, 16]. The doses of PMSG and hCG injected to each strain were optimized by our preliminary experiments.…”

Section: Methodsmentioning

confidence: 99%

<i>In vitro</i> maturation of immature rat oocytes under simple culture conditions and subsequent developmental ability

Taketsuru

Kaneko

2016

Journal of Reproduction and Development

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Rat oocytes can be produced artificially by superovulation. Because some strains show low sensitivity to superovulation treatment, in vitro maturation is an alternative method to produce numerous matured oocytes. Furthermore, establishment of an in vitro maturation system with simple culture conditions is cost effective and leads to easy handling of oocytes. This study examined developmental ability of rat germinal vesicle (GV) oocytes maturing in vitro under simple culture conditions. Significantly different numbers of ovulated oocytes reached the second metaphase of meiosis (MII) among Jcl:Wistar (17.0), F344/Stm (31.0), and BN/SsNSlc (2.2) rats in whom superovulation was induced by pregnant mare serum gonadotropin (PMSG) and human chorionic gonadotropin. However, similar numbers of GV oocytes were obtained from ovaries of PMSG-injected Wistar (27.7), F344 (34.7), and BN (24.7) rats. These GV oocytes were cultured in vitro in HTF, αMEM, and a 1:1 HTF + αMEM or TYH + αMEM mixture. High proportions of Wistar and F344 oocytes that matured to MII in αMEM were parthenogenetically activated by strontium chloride treatment (78% and 74%, respectively). Additionally, 10% of matured oocytes of both strains developed into offspring after intracytoplasmic sperm injection and embryo transfer to foster mothers. Although BN oocytes cultured in αMEM could be parthenogenetically activated and developed into offspring, the success rate was lower than that for Wistar and F344 oocytes. This study demonstrated that numerous GV oocytes were produced in rat ovaries by PMSG injection. This simple in vitro maturation system of immature oocytes could be further developed to maintain valuable rat strains experiencing reproductive difficulties.

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“…embryo cryopreservation. The 2-cell embryos were frozen via the vitrification method 18 . The embryos were placed in a solution of 10% propylene glycol in PB1 26 at room temperature for 10 min and then transferred in 5 μL aliquots into 1.2 mL serum tubes (Sumitomo Bakelite Co. Ltd, Tokyo, Japan).…”

Section: Methodsmentioning

confidence: 99%

“…Various strains of the human disease model have been rapidly produced by simple endonuclease introduction technique into embryos using microinjection [12][13][14] and the electroporation (TAKE -Technique for Animal Knockout system by Electroporation) method [15][16][17] . ET is required for the efficient production of new strains from these genome-edited embryos and the regeneration of valuable strains from frozen embryos that are used as genetic resources [18][19][20] .…”

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confidence: 99%

Simple induction of pseudopregnancy by artificial stimulation using a sonic vibration in rats

Kaneko

Endo

Tsunoda

et al. 2020

Sci Rep

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embryo transfer has been used as one of the essential reproductive technologies for production of new strains and maintenance of genetic resources in animals. Mating with vasectomised male rats is a requirement for inducing pseudopregnancy in female rats selected for embryo transfer. Although this procedure has been used routinely, large breeding space and high expenditure are required to maintain a sufficient number of females and vasectomised males. This study was performed to induce pseudopregnancy in females by artificial stimulation using sonic vibration instead of vasectomised males. The females continued to be in the dioestrus stage for at least 14 days after artificial stimulation was performed. Of fresh 2-cell embryos that transferred into the oviducts of females after artificial stimulation, 56% was implanted and 50% was developed to offspring. Approximately 46% of the frozen 2-cell embryos were implanted and 24% developed into offspring. Furthermore, 66% of the fresh pronuclear embryos were implanted and 60% developed into offspring. This study successfully induced pseudopregnancy in rat females by artificial stimulation using a sonic vibration. This method, 'Easy-ET', was useful for efficient production and maintenance of rat strains.

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Efficient collection and cryopreservation of embryos in F344 strain inbred rats

Cited by 26 publications

References 24 publications

Allele-specific genome editing and correction of disease-associated phenotypes in rats using the CRISPR–Cas platform

Allele-specific genome editing and correction of disease-associated phenotypes in rats using the CRISPR–Cas platform

<i>In vitro</i> maturation of immature rat oocytes under simple culture conditions and subsequent developmental ability

Simple induction of pseudopregnancy by artificial stimulation using a sonic vibration in rats

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