Production of somatic cell nuclear transfer embryos using <i>in vitro</i>-grown and <i>in vitro</i>-matured oocytes in rabbits

Sugimoto, Hironobu; Kida, Yutaka; Oh, Noriyoshi; Kitada, Kensaku; Matsumoto, Kazuya; Saeki, Kazuhiro; Taniguchi, Takeshi; Hosoi, Yoshihiko

doi:10.1017/s0967199414000082

Cited by 6 publications

(7 citation statements)

References 39 publications

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“…Animal somatic cell nuclear transfer (SCNT) technology has greatly expanded the fields of life science and biotechnology (Jin, Lee, Taweechaipaisankul, Kim, & Lee, ; Sugimoto et al, ), although aberrant reprogramming of epigenetic modifications from the donor cell genome still hampers the efficiency of this procedure (Hua et al, ; Niemann, Tian, King, & Lee, ). Donor cell nuclei possess specific epigenetic information, encoded as histone modifications and DNA methylation (Hochedlinger & Jaenisch, ), that can affect early SCNT embryo development.…”

Section: Introductionmentioning

confidence: 99%

Epigenetic states of donor cells significantly affect the development of somatic cell nuclear transfer (SCNT) embryos in pigs

Zhai

Zhang

et al. 2017

Molecular Reproduction Devel

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The type and pattern of epigenetic modification in donor cells can significantly affect the developmental competency of somatic cell nuclear transfer (SCNT) embryos. Here, we investigated the developmental capacity, gene expression, and epigenetic modifications of SCNT embryos derived from porcine bone marrow-derived mesenchymal stem cells (BMSCs) and fetal fibroblasts (FFs) donor cells compared to embryos obtained from in vitro fertilization (IVF). Compared to FFs, the donor BMSCs had more active epigenetic markers (Histone H3 modifications: H3K9Ac, H3K4me3, and H3K4me2) and fewer repressive epigenetic markers (H3K9me3, H3K9me2, and DNA methyltransferase 1). Embryos derived from BMSC nuclear-transfer (BMSC-NT embryos) and IVF embryos had significantly higher cleavage and blastocyst rates (BMSC-NT: 71.3 ± 3.4%, 29.1 ± 2.3%; IVF: 69.2 ± 2.2%, 30.2 ± 3.3%; respectively) than FF-NT embryos (58.1 ± 3.4%, 15.1 ± 1.5%, respectively). Bisulfite sequencing revealed that DNA methylation at the promoter regions of NANOG and POU5F1 was lower in BMSC-NT embryos (30.0%, 9.8%, respectively) than those in FF-NT embryos (34.2%, 28.0%, respectively). We also found that BMSC-NT embryos had more H3K9Ac and less H3K9me3 and 5-methylcytosine than FF-NT embryos. In conclusion, our finding comparing BMSCs versus FFs as donors for nuclear transfer revealed that differences in the initial epigenetic state of donor cells have a remarkable effect on overall nuclear reprogramming of SCNT embryos, wherein donor cells possessing a more open chromatin state are more conducive to nuclear reprogramming.

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Section: Introductionmentioning

confidence: 99%

Epigenetic states of donor cells significantly affect the development of somatic cell nuclear transfer (SCNT) embryos in pigs

Zhai

Zhang

et al. 2017

Molecular Reproduction Devel

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“…Somatic cell nuclear transfer (SCNT) is an important technique for producing genome-edited animals; it also has great value in saving endangered species and in clone stem cell therapy, and the rabbit is an important model for studying SCNT (Liu et al ., 2016; Matoba & Zhang, 2018). However, SCNT has a low efficiency (Sugimoto et al ., 2015). Many factors, such as the type and cell cycle of the donor cell, the oocyte status, the fusion method, and the culture medium, affect the success of cloning (Matoba & Zhang, 2018).…”

Section: Introductionmentioning

confidence: 99%

“…However, abnormal reprogramming is the main reason for the low efficiency of SCNT embryos (Chen et al ., 2013). Consequently, as the proteins, RNAs, and other factors in oocytes play a pivotal role in reprogramming and development, the selection of suitable oocytes is crucial to the success of SCNT (Sugimoto et al ., 2015).…”

Section: Introductionmentioning

confidence: 99%

Use of oocytes selected by brilliant cresyl blue staining enhances rabbit cloned embryo development in vitro

Jia

Ding

Shen

et al. 2019

Zygote

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SummaryRabbits play an important role in people’s lives due to their high nutritional value and high-quality hair that can be used as raw material for textiles. Furthermore, rabbits are an important animal model for human disease, as genome-edited animals are particularly valuable for studying gene functions and pathogenesis. Somatic cell nuclear transfer (SCNT) is an important technique for producing genome-edited animals and it has great value in saving endangered species and in clone stem cell therapy. However, the low efficiency of SCNT limits its application, with the selection of suitable rabbit oocytes being crucial to its success. In the present study, we collected oocytes from ovarian follicles and stained them with 26 μM brilliant cresyl blue (BCB). We then matured the oocytes in vitro and used them for SCNT. Comparison of the BCB-positive oocytes with BCB-negative oocytes and the control group showed that the BCB-positive group had a significantly higher maturation rate (81.4% vs. 48.9% and 65.3% for the negative and control groups, respectively), cleavage rate (86.6% vs. 67.9% and 77.9%), blastocyst rate (30.5% vs. 12.8% and 19.6%), total number of blastocysts (90±7.5 vs. 65.3±6.3 and 67.5±5.7), and inner cell mass (ICM)/ trophectoderm (TE) index (42.3±4.2 vs. 30.2±2.1 and 33.9±5.1) (P<0.05). The BCB-positive group had a significantly lower apoptosis index (2.1±0.6 vs. 8.2±0.9 and 6.7±1.1 for the negative and control groups, respectively) (P<0.05). These findings demonstrate that BCB-positive oocytes have a higher maturation ability and developmental competence in vitro, indicating that BCB staining is a reliable method for selecting oocytes to enhance the efficiency of SCNT.

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“…Moreover, Tian J et al (2012) used EBSS complete medium for SCNT embryo culture and obtained good blastocyst rates ranging from 39 to 79% [ 151 ]; although these variations in outcome appeared to be mainly due to the donor nuclei used during SCNT. In rabbits, SCNT embryos were recently reconstructed after in vitro growth (IVG) and IVM of follicles, and subsequently cultured in supplemented CMRL-1066 medium [ 152 ]. Although oocytes grew and matured in vitro, their cleavage and blastocyst rates after fusion and activation were similar to those matured in vivo [ 152 ] .…”

Section: Introductionmentioning

confidence: 99%

“…In rabbits, SCNT embryos were recently reconstructed after in vitro growth (IVG) and IVM of follicles, and subsequently cultured in supplemented CMRL-1066 medium [ 152 ]. Although oocytes grew and matured in vitro, their cleavage and blastocyst rates after fusion and activation were similar to those matured in vivo [ 152 ] . This innovative result suggests that IVG-IVM oocytes have developmental competency and the resulting SCNT embryos were reprogrammed and supported by the medium in which they were cultured.…”

Section: Introductionmentioning

confidence: 99%

Choosing a culture medium for SCNT and iSCNT reconstructed embryos: from domestic to wildlife species

Cordova¹,

King²,

Mastromonaco³

2017

J Anim Sci Technol

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Over the past decades, in vitro culture media have been developed to successfully support IVF embryo growth in a variety of species. Advanced reproductive technologies, such as somatic cell nuclear transfer (SCNT), challenge us with a new type of embryo, with special nutritional requirements and altered physiology under in vitro conditions. Numerous studies have successfully reconstructed cloned embryos of domestic animals for biomedical research and livestock production. However, studies evaluating suitable culture conditions for SCNT embryos in wildlife species are scarce (for both intra- and interspecies SCNT). Most of the existing studies derive from previous IVF work done in conventional domestic species. Extrapolation to non-domestic species presents significant challenges since we lack information on reproductive processes and embryo development in most wildlife species. Given the challenges in adapting culture media and conditions from IVF to SCNT embryos, developmental competence of SCNT embryos remains low. This review summarizes research efforts to tailor culture media to SCNT embryos and explore the different outcomes in diverse species. It will also consider how these culture media protocols have been extrapolated to wildlife species, most particularly using SCNT as a cutting-edge technical resource to assist in the preservation of endangered species.

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Production of somatic cell nuclear transfer embryos using in vitro-grown and in vitro-matured oocytes in rabbits

Cited by 6 publications

References 39 publications

Epigenetic states of donor cells significantly affect the development of somatic cell nuclear transfer (SCNT) embryos in pigs

Epigenetic states of donor cells significantly affect the development of somatic cell nuclear transfer (SCNT) embryos in pigs

Use of oocytes selected by brilliant cresyl blue staining enhances rabbit cloned embryo development in vitro

Choosing a culture medium for SCNT and iSCNT reconstructed embryos: from domestic to wildlife species

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