Phytohemagglutinin improves efficiency of electrofusing mammary gland epithelial cells into oocytes in goats

Zhang, Y.L.; Liu, F.J.; Sun, Dongjun; Chen, X.Q.; Zhang, Y.; Zheng, Ying; Zhao, Minghui; Wang, G.H.

doi:10.1016/j.theriogenology.2007.10.028

Cited by 11 publications

(4 citation statements)

References 20 publications

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“…For caprine NT, which is a popular model, many somatic cell types, including those from mammary epithelia (Zhang et al. ; Yuan et al. ), ovarian cumulus (Zou et al.…”

Section: Introductionmentioning

confidence: 99%

“…One of the key factors is the nuclear donor cell type. For caprine NT, which is a popular model, many somatic cell types, including those from mammary epithelia (Zhang et al 2008;Yuan et al 2009), ovarian cumulus (Zou et al 2001;Keefer et al 2002), foetal fibroblasts (Baguisi et al 1999;Keefer et al 2002;Melican et al 2005;Soh et al 2012) and skin (Wang et al 2007;Abdullah et al 2011;Kwong et al 2012;Wan et al 2012), have been utilized. A clear consensus has not yet been reached as to which somatic cell type is best for nuclear transfer.…”

Section: Introductionmentioning

confidence: 99%

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Comparison of In Vitro Developmental Competence of Cloned Caprine Embryos Using Donor Karyoplasts from Adult Bone Marrow Mesenchymal Stem Cells vs Ear Fibroblast Cells

Kwong

Nam

Khadijah

et al. 2014

Reprod Domestic Animals

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The aim of this study was to produce cloned caprine embryos using either caprine bone marrow-derived mesenchymal stem cells (MSCs) or ear fibroblast cells (EFCs) as donor karyoplasts. Caprine MSCs were isolated from male Boer goats of an average age of 1.5 years. To determine the pluripotency of MSCs, the cells were induced to differentiate into osteocytes, chondrocytes and adipocytes. Subsequently, MSCs were characterized through cell surface antigen profiles using specific markers, prior to their use as donor karyoplasts for nuclear transfer. No significant difference (p > 0.05) in fusion rates was observed between MSCs (87.7%) and EFCs (91.3%) used as donor karyoplasts. The cleavage rate of cloned embryos derived with MSCs (87.0%) was similar (p > 0.05) to those cloned using EFCs (84.4%). However, the in vitro development of MSCs-derived cloned embryos (25.3%) to the blastocyst stage was significantly higher (p < 0.05) than those derived with EFCs (20.6%). In conclusion, MSCs could be reprogrammed by caprine oocytes, and production of cloned caprine embryos with MSCs improved their in vitro developmental competence, but not in their fusion and cleavage rate as compared to cloning using somatic cells such as EFCs.

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“…For caprine NT, which is a popular model, many somatic cell types, including those from mammary epithelia (Zhang et al. ; Yuan et al. ), ovarian cumulus (Zou et al.…”

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Comparison of In Vitro Developmental Competence of Cloned Caprine Embryos Using Donor Karyoplasts from Adult Bone Marrow Mesenchymal Stem Cells vs Ear Fibroblast Cells

Kwong

Nam

Khadijah

et al. 2014

Reprod Domestic Animals

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“…The SCNT procedures were performed as described previously by our laboratory [20], [21]. Briefly, oocytes with the first polar body were selected for enucleation after 22–24 h of in vitro maturation.…”

Section: Methodsmentioning

confidence: 99%

Anti-Bacterial Activity of Recombinant Human β-Defensin-3 Secreted in the Milk of Transgenic Goats Produced by Somatic Cell Nuclear Transfer

Luo

et al. 2013

PLoS ONE

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The present study was conducted to determine whether recombinant human β-defensin-3 (rHBD3) in the milk of transgenic goats has an anti-bacterial activity against Escherichia coli (E. coli), Staphylococcus aureus (S. aureus) and Streptococcus agalactiae (S. agalactiae) that could cause mastitis. A HBD3 mammary-specific expression vector was transfected by electroporation into goat fetal fibroblasts which were used to produce fourteen healthy transgenic goats by somatic cell nuclear transfer. The expression level of rHBD3 in the milk of the six transgenic goats ranged from 98 to 121 µg/ml at 15 days of lactation, and was maintained at 90–111 µg/ml during the following 2 months. Milk samples from transgenic goats showed an obvious inhibitory activity against E. coli, S. aureus and S. agalactiae in vitro. The minimal inhibitory concentrations of rHBD3 in milk against E. coli, S. aureus and S. agalactiae were 9.5–10.5, 21.8–23.0 and 17.3–18.5 µg/mL, respectively, which was similar to those of the HBD3 standard (P>0.05). The in vivo anti-bacterial activities of rHBD3 in milk were examined by intramammary infusion of viable bacterial inoculums. We observed that 9/10 and 8/10 glands of non-transgenic goats infused with S. aureus and E. coli became infected. The mean numbers of viable bacteria went up to 2.9×103 and 95.4×103 CFU/ml at 48 h after infusion, respectively; the mean somatic cell counts (SCC) in infected glands reached up to 260.4×105 and 622.2×105 cells/ml, which were significantly higher than the SCC in uninfected goat glands. In contrast, no bacteria was presented in glands of transgenic goats and PBS-infused controls, and the SSC did not significantly change throughout the period. Moreover, the compositions and protein profiles of milk from transgenic and non-transgenic goats were identical. The present study demonstrated that HBD3 were an effective anti-bacterial protein to enhance the mastitis resistance of dairy animals.

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“…In SCNT, the fusion of somatic cells and enucleated oocytes usually uses electrofusion. Experimental studies have been conducted on many animals, such as pigs 143 - 154 , cattle 146 - 150 , goats 151 , 152 , and mice 153 - 155 . The specific research content and conclusions are summarized in Table 5 .…”

Section: Oocyte Electrical Activation/reconstructed Embryo Electrofusionmentioning

confidence: 99%

Recent advances in critical nodes of embryo engineering technology

Ma¹,

Gu²,

Chen³

et al. 2021

Theranostics

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The normal development and maturation of oocytes and sperm, the formation of fertilized ova, the implantation of early embryos, and the growth and development of foetuses are the biological basis of mammalian reproduction. Therefore, research on oocytes has always occupied a very important position in the life sciences and reproductive medicine fields. Various embryo engineering technologies for oocytes, early embryo formation and subsequent developmental stages and different target sites, such as gene editing, intracytoplasmic sperm injection (ICSI), preimplantation genetic diagnosis (PGD), and somatic cell nuclear transfer (SCNT) technologies, have all been established and widely used in industrialization. However, as research continues to deepen and target species become more advanced, embryo engineering technology has also been developing in a more complex and sophisticated direction. At the same time, the success rate also shows a declining trend, resulting in an extension of the research and development cycle and rising costs. By studying the existing embryo engineering technology process, we discovered three critical nodes that have the greatest impact on the development of oocytes and early embryos, namely, oocyte micromanipulation, oocyte electrical activation/reconstructed embryo electrofusion, and the in vitro culture of early embryos. This article mainly demonstrates the efforts made by researchers in the relevant technologies of these three critical nodes from an engineering perspective, analyses the shortcomings of the current technology, and proposes a plan and prospects for the development of embryo engineering technology in the future.

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Phytohemagglutinin improves efficiency of electrofusing mammary gland epithelial cells into oocytes in goats

Cited by 11 publications

References 20 publications

Comparison of In Vitro Developmental Competence of Cloned Caprine Embryos Using Donor Karyoplasts from Adult Bone Marrow Mesenchymal Stem Cells vs Ear Fibroblast Cells

Comparison of In Vitro Developmental Competence of Cloned Caprine Embryos Using Donor Karyoplasts from Adult Bone Marrow Mesenchymal Stem Cells vs Ear Fibroblast Cells

Anti-Bacterial Activity of Recombinant Human β-Defensin-3 Secreted in the Milk of Transgenic Goats Produced by Somatic Cell Nuclear Transfer

Recent advances in critical nodes of embryo engineering technology

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