Early Development of Cloned Bovine Embryos Produced from Oocytes Enucleated by Fluorescence Metaphase II Imaging Using a Conventional Halogen-Lamp Microscope

Iwamoto, D.; Yamagata, Kazuya; Kishi, Michio J.; Hayashi‐Takanaka, Yoko; Kimurâ, Hiroshi; Wakayama, Teruhiko; Saeki, Kazuhiro

doi:10.1089/cell.2014.0086

Cited by 5 publications

(5 citation statements)

References 32 publications

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“…In cattle, although the blastocyst production is similar to that obtained in in vitro production of embryos (IVEP), (Iwamoto et al, 2015;Song et al, 2015;Jeong et al, 2013;Kato et al, 2000), a high rate of fetal loss is observed at the beginning of pregnancy (Keefer, 2015). These losses generally occur during placentation and have been related to epigenetic changes caused by failures in reprogramming of the differentiated donor cells (Campbell et al, 2007).…”

Section: Introductionmentioning

confidence: 94%

Meiotic arrest as an alternative to increase the production of bovine embryos by somatic cell nuclear transfer

Caixeta

Sousa

Guimarães

et al. 2016

Zygote

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This study aimed to evaluate the effect of meiotic arrest using phosphodiesterase type 3A (PDE 3A) inhibitors, cilostamide and C-type natriuretic peptide (NPPC), on pre-maturation (PM) of oocytes to be used in the production of cloned embryos. Nuclear maturation, in vitro embryo production (IVP), somatic cell nuclear transfer (SCNT) and parthenogenetic activation (PA), and total cells number of cloned embryos were evaluated. The results were analysed by chi-squared and Kruskal-Wallis test with a P-value 0.05) between control and PM, both for cleavage (78.2% and 76.9%) and blastocyst (35.5% and 29.3%) rates. After SCNT, cleavage rate was also similar (P > 0.05) between control and PM (66% and 51.9%) however, blastocyst rate was lower (P< 0.05) in the PM group than in the control group (7.4% and 30.2%). After 6 h of PM with 100 nM of NPPC, approximately 84.9% of the oocytes remained at GV. No difference was found between control and PM in cleavage (69.2% and 76.1%) and blastocyst rates (37,4% and 35%) after IVP. Similarly, no differences between PM and control groups were observed for cleavage (69.2% and 68.4%) and blastocyst (24.4% and 21.5%) rates. SCNT and PA embryos from control or PM oocytes had similar total cell number. It can be concluded that PM for 6 h with 100 nM NPPC is feasible for cloned embryo production without affecting embryo outcome.

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Section: Introductionmentioning

confidence: 94%

Meiotic arrest as an alternative to increase the production of bovine embryos by somatic cell nuclear transfer

Caixeta

Sousa

Guimarães

et al. 2016

Zygote

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“…Hence, a more recent method introduces fluorescence-labelled antibodies, which can be injected into the oocytes. These antibodies will bind to chromosomes and are visible under a halogen-lamp microscope [12].…”

Section: Removal Of Zona Pellucida and Enucleationmentioning

confidence: 99%

Handmade cloning: a developing alternative to traditional SCNT cloning methods

Chen

2022

International Conference on Biomedical and Intelligent Systems (IC-BIS 2022)

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Handmade Cloning (HMC) is a method of somatic cell nuclear transfer (SCNT) that eliminates the need for micromanipulators, making it a novel revolution in embryology. Not only is it cheaper than its predecessor, traditional cloning that uses micromanipulators, but it is also more efficient in terms of birth rate and economic expenditures. Given that the technology is relatively recent, variations in HMC procedure and method have been explored in different species and experiments; these modifications generally have a positive effect on HMC efficiency in such experiments. In recent years, the increasing number of research in HMC generally fall into two categories: first, the factors that contribute to HMC’s improved efficiency. Two, HMC’s possible usage in genealogy and transgene integration. Hence, this review will highlight and summarize the factors that contribute to HMC efficiency, the issues and setbacks that obstruct HMC development, as well as a glimpse into the future of HMC.

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“…Moreover, lipid droplets in the ooplasm obstruct identification and enucleation of metaphase II (MII) chromosomes, especially in bovine species, ultimately affecting the efficiency of the process. To overcome this problem, a new experimental system have been employed, which is based on fluorescent observation of chromosomes in living oocytes without any damage by injecting fluorescence-labeled antibody conjugates that bind to chromosomes and fluorescent observation using a conventional halogen-lamp microscope [ 34 ].…”

Section: Step By Step Guide To Hmcmentioning

confidence: 99%

Handmade cloning: recent advances, potential and pitfalls

Verma

Arora

Sethi

et al. 2015

J Animal Sci Biotechnol

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Handmade cloning (HMC) is the most awaited, simple and micromanipulator-free version of somatic cell nuclear transfer (SCNT). The requirement of expensive micromanipulators and skilled expertise is eliminated in this technique, proving it as a major revolution in the field of embryology. During the past years, many modifications have been incorporated in this technique to boost its efficiency. This alternative approach to micromanipulator based traditional cloning (TC) works wonder in generating comparable or even higher birth rates in addition to declining costs drastically and enabling cryopreservation. This technique is not only applicable to intraspecies nuclear transfer but also to interspecies nuclear transfer (iSCNT) thus permitting conservation of endangered species. It also offers unique possibilities for automation of SCNT which aims at production of transgenic animals that can cure certain human diseases by producing therapeutics hence, providing a healthier future for the wellbeing of humans. The present review aims at highlighting certain aspects of HMC including recent advancements in procedure and factors involved in elevating its efficiency besides covering the potentials and pitfalls of this technique.

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Early Development of Cloned Bovine Embryos Produced from Oocytes Enucleated by Fluorescence Metaphase II Imaging Using a Conventional Halogen-Lamp Microscope

Cited by 5 publications

References 32 publications

Meiotic arrest as an alternative to increase the production of bovine embryos by somatic cell nuclear transfer

Meiotic arrest as an alternative to increase the production of bovine embryos by somatic cell nuclear transfer

Handmade cloning: a developing alternative to traditional SCNT cloning methods

Handmade cloning: recent advances, potential and pitfalls

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