• Title/Summary/Keyword: Pig embryo

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In Vitro Production of Pig Embryos using Intracytoplasmic Injection of Flow Cytometry Sorted Boar Spermatozoa

  • Kim, Dae-Young;Hyun, Sang-Hwan;Lee, Eun-Song
    • Journal of Embryo Transfer
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    • v.23 no.4
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    • pp.275-281
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    • 2008
  • The ability to preselect the sex of piglets is advantageous in the pig industry. The objective of this study was to examine the feasibility of using intracytoplasmic sperm injection (ICSI) with sorted spermatozoa to produce piglets with a preselected sex. Pig embryos were produced by ICSI of frozen X- and Y-sperm that had been separated by flow cytometry. The developmental competence of the embryos was investigated in vitro and in vivo. The populations of X- and Y-spermatozoa were 52.7% and 47.3%, respectively in our samples. The in vitro development of ICSI embryos was enhanced by longer of in vitro maturation of oocytes ($44{\sim}48\;h$ vs. $40{\sim}43\;h$). Their cleavage ($65{\sim}70%$) and blastocyst formation ($9{\sim}12%$) rates were not significantly different between male and female ICSI embryos, or between sorted and unsorted sperm-derived embryos. One pregnancy was established in a recipient that was transferred with 110 female ICSI embryos, but the pregnancy was terminated on Day 89 of gestation. Our results suggest that the separation X- and Y-spermatozoa by flow cytometric sorting can be a useful tool in combination with ICSI for the production of pig embryos and piglets of preselected sex.

Factors Influencing the Efficiency of In Vitro Embryo Production in the Pig

  • Lin, Tao;Lee, Jae Eun;Shin, Hyun Young;Oqani, Reza K.;Jin, Dong Il
    • Reproductive and Developmental Biology
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    • v.39 no.2
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    • pp.29-36
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    • 2015
  • Pigs are considered an ideal source of human disease model due to their physiological similarities to humans. However, the low efficiency of in vitro embryo production (IVP) is still a major barrier in the production of pig offspring with gene manipulation. Despite ongoing advances in the associated technologies, the developmental capacity of IVP pig embryos is still lower than that of their in vivo counterparts, as well as IVP embryos of other species (e.g., cattle and mice). The efficiency of IVP can be influenced by many factors that affect various critical steps in the process. The previous relevant reviews have focused on the in vitro maturation system, in vitro culture conditions, in vitro fertilization medium, issues with polyspermy, the utilized technologies, etc. In this review, we concentrate on factors that have not been fully detailed in prior reviews, such as the oocyte morphology, oocyte recovery methods, denuding procedures, first polar body morphology and embryo quality.

Microscopic Study of the Pig Peri-implantation Embryos (전자현미경에 의한 착상 전후 돼지수정란의 형태학적 변화에 관한 연구)

  • 김진회;백청순;이훈택;정길생
    • Korean Journal of Animal Reproduction
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    • v.18 no.2
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    • pp.141-150
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    • 1994
  • Morphological features of the interaction between the hatching blastocyst and implantation in pig were studied by electron microscopy. The observations extended from late blastocyst stage to the completion of trophoblastic erosion of the epithelium and early decidual transformation of the epithelium and early decidual transformation of the stromal cells. Between day 7 and 17 of pregnancy, blastocysts from 0.3 to 12 mm in diameter were flushed from the uterine horns of Dutch Landrace pigs. On the 7th of development in the pig blastocyst, the blastocyst shedded of the zona pellucida established the tips of microvilli and with bleb-like cytoplasmic protrusions of the epithelial cells. From day 11 on in pig embryo, the bilayered trophoblast undergoes a dramatic phase of elongation so that the initially spherical expanded blastocyst becomes tubular. In pig, close apposition to the uterine wall beg-ins at about 12 $^1$/$_2$ days and then attachment occurred during the afternoon of the 16th or 18th day post coitum. At this stage, embryonic loss compared with corpus luteum number is up to 40% of ovulated oocytes. Therefore, the implantation failture of these embryos may be mainly caused by morphological abnormality and failture of zona shedding.

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Selection of Early Cleaved Embryos and Optimal Recipients to Improve Efficiency of Pig Cloning

  • Koo, Ok-Jae;Lee, Dong-Won;Kang, Jung-Taek;Kwon, Dae-Kee;Park, Hee-Jung;Park, Sol-Ji;Kim, Su-Jin;Jang, Goo;Lee, Byeong-Chun
    • Journal of Embryo Transfer
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    • v.25 no.4
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    • pp.221-227
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    • 2010
  • Early cleavage is a reliable prognostic tool for successful embryo transfer in assisted reproduction because early cleaved embryo show better pregnancy rate after transfer. There for, preparation of good embryo recipient is important factor to optimize efficiency of pig cloning. The present study was performed to evaluate the effect of early cleavage on the in vivo development of cloned embryos and to analyze breed, parity and estrous synchrony to optimize recipient for pig cloning. In vitro matured porcine oocytes derived from local slaughterhouse and fibroblasts derived from miniature pig fetuses were used for somatic cell nuclear transfer (SCNT). Reconstructed embryos were transferred to recipient pigs on the same day of SCNT or after 1~2 days of in vitro culture for selecting early cleaved embryos. Breed, parity and date of standing estrous of recipients were recorded for analysis. After 25~35 days after embryo transfer pregnancy was diagnosed using ultrasonography, and pregnant recipients were monitored till delivery. Between purebred and crossbred, no significant difference was founded in both pregnancy and delivery rates. However, early cleaved embryos showed significantly higher pregnancy (46.2%) and delivery (12.8%) rates compared to non-selectively transferred group (24.8% and 4.5%, respectively). The results also showed that the recipients showing standing estrous on the same day of SCNT and less than 4 parities were most suitable for pig cloning.

The Present Situation and Problems of In Vitro Fertilization in Swine (돼지 체외수정의 현황과 문제점)

  • 류일선
    • Journal of Embryo Transfer
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    • v.7 no.1
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    • pp.41-47
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    • 1992
  • 1. In vitro system, LR and FSR accelerated and facilitated meiotic progression, and LH selectively improved cytoplasmic maturation which is required to promote the formation of a male pronucleus. 2. Caffeine (2mM) in the fetilization medium was required not only for inducing zona penetrating ability of boar also for developing to the male pronucleus of the penetrat- ing spermatozoa in vitro. 3. The germinal vesicle (GV)stage was observed for the first 17.6 hr;germinal vesicle break-down (GVBD)stage between 17.6~26.4 hr ;metaphase I (M-I)from 26.4 - 30. 9hr;anaphase I(A-I)ranged from 30. 9~33.4hr;telophase I(T-I) at 33.4~34.4hr; and metaphase II(M-II) at 34.4-48hr. 4. The addition of 10%(v /v) pig follicular fluid (pFF) to maturation media significantly increased the rate of nuclear maturation of pig oocytes (p<0.01), whereas the rate of nuclear maturation of pig oocytes among three different media did not differ. 5. The presence of a primary culture of POEC promotes in vitro development of early cleavage stage pig embryos.

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Studies on Embryo Transfer in Pigs (돼지의 수정란이식에 관한 고찰)

  • 손동수
    • Journal of Embryo Transfer
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    • v.3 no.1
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    • pp.6-12
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    • 1988
  • The possibility of embryo transfer technique in pigs to introduce a new genetic material into closed herds for disease control was investigated. The results investigated were as follows: 1. The exhibiting rate of estrus on the administration of altrenogest and PMSG ranged from 83.3 to 100% and the estrus exhibited within 4.0 - 5.3 days after the administration of altrenogest. 2. The average number of ovulation points per pig by the injection of PMSG and HCG were 14.0 - 30.7. 3. The average number of recovered embryos per pig was 15.7, and 72.8% of embryos were recovered. 4. The pregnancy rate of recipients was 63.6% and the survival rate of transferred embryos were 21%. 5. When the 1-cell embryos were cultured for 24hrs, they were all developed. Therefore it is possible to recover and preserve the embryos from the donor pigs which have high genetic ability, and to transfer embryos to recipient pigs which are separated from donor pigs in Korea.

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