Pregnancies and piglets from large white sow recipients after two transfer methods of cloned and transgenic embryos of different pig breeds

Schmidt, M.; Kragh, P. M.; Li, J.; Du, Yutao; Lin, Liang; Liu, Y.; Bøgh, Ingrid Brück; Winther, K.D.; Vajta, Gábor; Callesen, Henrik

doi:10.1016/j.theriogenology.2010.05.026

Cited by 63 publications

(97 citation statements)

References 33 publications

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“…This suggests that embryos generated from fibroblasts of Chinese local LW pigs rather than Western pig breeds allow their surrogate mothers to establish pregnancy and maintain pregnancy to term more easily. However, this seems contrary to the finding that pregnancy rate and delivery rate were significantly increased if the transferred cloned embryos and their recipient are of the same breed (Koo et al, 2009;Schmidt et al, 2010). In the present study, LY or YL hybrid sows were used as recipients, which are genetically more similar with cloned embryos generated from P,D,L,YFFBs and P,D,L,Y-AFBs.…”

Section: Discussioncontrasting

confidence: 66%

“…This negative effect could be minimized by increasing the number of transferred cloned embryos to an individual surrogate. Schmidt et al (2010) showed that when the number of transferred cloned blastocysts selected with normal morphology was increased from < 60 to 60-120 per recipient, the surrogates' pregnancy rate and delivery rate were significantly elevated. However, in our study, the cloned embryos were unselected and transferred at around two-cell stage.…”

Section: Discussionmentioning

confidence: 99%

“…Success in production of cloned pigs from somatic cells was first reported almost at the same time in 2000 by three independent groups (Betthauser et al, 2000;Onishi et al, 2000;Polejaeva et al, 2000). Since then, the somatic cell nuclear transfer (SCNT) technique has been used to amplify superior pigs or has been applied in combination with genetic modifications to generate different types of transgenic or knockout pigs (Klymiuk et al, 2010;Prather et al, 2008;Schmidt et al, 2010;Vajta and Callesen 2012). Even though the pig SCNT procedure has been developed for over 10 years and a large number of cloned pigs have been produced, this approach is still inefficient.…”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Effects of Donor Fibroblast Cell Type and Transferred Cloned Embryo Number on the Efficiency of Pig Cloning

Shi

Liu

et al. 2013

Cellular Reprogramming

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Currently, cloning efficiency in pigs is very low. Donor cell type and number of cloned embryos transferred to an individual surrogate are two major factors that affect the successful rate of somatic cell nuclear transfer (SCNT) in pigs. This study aimed to compare the influence of different donor fibroblast cell types and different transferred embryo numbers on recipients' pregnancy rate and delivery rate, the average number of total clones born, clones born alive and clones born healthy per litter, and the birth rate of healthy clones ( = total number of healthy cloned piglets born / total number of transferred cloned embryos). Three types of donor fibroblasts were tested in large-scale production of cloned pigs, including fetal fibroblasts (FFBs) from four genetically similar Western swine breeds of Pietrain (P), Duroc (D), Landrace (L), and Yorkshire (Y), which are referred to as P,D,LY-FFBs, adult fibroblasts (AFBs) from the same four breeds, which are designated P,D,L,Y-AFBs, and AFBs from a Chinese pig breed of Laiwu (LW), which is referred to as LW-AFBs. Within each donor fibroblast cell type group, five transferred cloned embryo number groups were tested. In each embryo number group, 150-199, 200-249, 250-299, 300-349, or 350-450 cloned embryos were transferred to each individual recipient sow. For the entire experiment, 92,005 cloned embryos were generated from nearly 115,000 matured oocytes and transferred to 328 recipients; in total, 488 cloned piglets were produced. The results showed that the mean clones born healthy per litter resulted from transfer of embryos cloned from LW-AFBs (2.53 -0.34) was similar with that associated with P,D,L,Y-FFBs (2.72 -0.29), but was significantly higher than that resulted from P,D,L,Y-AFBs (1.47 -0.18). Use of LW-AFBs as donor cells for SCNT resulted in a significantly higher pregnancy rate (72.00% vs. 59.30% and 48.11%) and delivery rate (60.00% vs. 45.93% and 35.85%) for cloned embryo recipients, and a significantly higher birth rate of healthy clones (0.5009% vs. 0.3362% and 0.2433%) than that resulting from P,D,L,Y-AFBs and P,D,L,Y-FFBs. This suggests that using LW-AFBs as donor cells results in a higher cloning efficiency in pigs, compared with the other two donor fibroblast cell types. The birth rate of healthy clones was significantly improved when the number of transferred cloned embryos was increased from 150-199 to 200-450 per recipient. However, increase of the number of transferred embryos from 200-249 to 250-450 per surrogate did not change the birth rate of healthy clones. This suggests that transfer of excessive (250-450) cloned embryos to an individual surrogate is not necessary for increasing the cloning efficiency in pigs, and the relatively optimal number of reconstructed embryos transferred to individual recipient is 200-249. Furthermore, our results indicated that the numbers of total born clones, clones born alive, and clones born healthy per litter have a significantly high positive correlation with each other. The present study provi...

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Section: Discussioncontrasting

confidence: 66%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Effects of Donor Fibroblast Cell Type and Transferred Cloned Embryo Number on the Efficiency of Pig Cloning

Shi

Liu

et al. 2013

Cellular Reprogramming

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“…Donor cells were from cultured ear fibroblasts obtained from a Danish Landrace · Yorkshire (65%:35%) sow. The cloned embryos were surgically transferred to surrogate sows 5-6 days after cloning as described in Schmidt et al (2010). Normal litters were used as controls (75% Danish Landrace:25% Yorkshire).…”

Section: Animals Diets and Samplingmentioning

confidence: 99%

Expression of Innate Immune Response Genes in Liver and Three Types of Adipose Tissue in Cloned Pigs

Rødgaard

Skovgaard

Stagsted

et al. 2012

Cellular Reprogramming

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The pig has been proposed as a relevant model for human obesity-induced inflammation, and cloning may improve the applicability of this model. We tested the assumptions that cloning would reduce interindividual variation in gene expression of innate immune factors and that their expression would remain unaffected by the cloning process. We investigated the expression of 40 innate immune factors by high-throughput quantitative real-time PCR in samples from liver, abdominal subcutaneous adipose tissue (SAT), visceral adipose tissue (VAT), and neck SAT in cloned pigs compared to normal outbred pigs.The variation in gene expression was found to be similar for the two groups, and the expression of a small number of genes was significantly affected by cloning. In the VAT and abdominal SAT, six out of seven significantly differentially expressed genes were downregulated in the clones. In contrast, most differently expressed genes in both liver and neck SAT were upregulated (seven out of eight). Remarkably, acute phase proteins (APPs) dominated the upregulated genes in the liver, whereas APP expression was either unchanged or downregulated in abdominal SAT and VAT. The general conclusion from this work is that cloning leads to subtle changes in specific subsets of innate immune genes. Such changes, even if minor, may have phenotypic effects over time, e.g., in models of long-term inflammation related to obesity.

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“…also to identify the good or bad donors. For this, donors were characterized through their patterns of progesterone or estradiol concentrations in plasma and milk (Callesen et al, 1988(Callesen et al, , 1990 together with detailed estrus observations (Callesen et al, 1993c), or by use of ultrasound examinations of follicular Anim. Reprod., v.13, n.3, p.334-339, Jul./Sept.…”

Section: The Superovulated Donor Cowmentioning

confidence: 99%

Circles around the farm animal embryo a Danish perspective

Callesen

2016

Anim Reprod

Self Cite

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With focus on the farm animal embryo, a short overview is given about my research activities over the last 35 years. These activities have been described in five circles, covering different key aspects of my work. The first circle included studies on the basic biology related to oocyte maturation and follicular endocrinology in superovulated dairy cows. Methods were developed to characterize the donor cows with respect to their production of transferable embryos, and some were implemented into a Danish MOET breeding plan. The second circle dealt with in-vitro embryo production in cattle with development of a protocol to produce such embryos at high and consistent levels. Several comparisons were made to reveal consequences of the artificial in-vitro methods on oocytes and embryos, but also through studies of the newborn calves. The third circle was related to development and implementation of a number of technologies within this broad field; examples are mentioned for both oocyte recovery from donor cows, different steps in the in-vitro embryo production system, new ways to perform vitrification and nuclear transfer, and finally a new system to determine oxygen consumption in single embryos. In the fourth circle is described activities from the last years, where work was done with focus on the pregnancy rates after insemination of the dairy cow in their post-partum period, and where somatic cell nuclear transfer was developed both as a technology in itself as well as a helping technique to produce transgenic pigs as models for important human diseases. The fifth and final circle is addressing and thanking the many colleagues and collaboration partners that I have been involved with during all the years to do this work. Nothing could or would have been the same without them and their participation.

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Pregnancies and piglets from large white sow recipients after two transfer methods of cloned and transgenic embryos of different pig breeds

Cited by 63 publications

References 33 publications

Effects of Donor Fibroblast Cell Type and Transferred Cloned Embryo Number on the Efficiency of Pig Cloning

Effects of Donor Fibroblast Cell Type and Transferred Cloned Embryo Number on the Efficiency of Pig Cloning

Expression of Innate Immune Response Genes in Liver and Three Types of Adipose Tissue in Cloned Pigs

Circles around the farm animal embryo a Danish perspective

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