• Journal of Embryo Transfer
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• v.31 no.2
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• pp.117-121
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• 2016

Although piglets have been delivered by embryo transfer (ET) with in vitro produced (IVP) embryos and blastocysts, a success rate has still remained lower level. Unlike mouse, human, and bovine, it is difficult to a production of piglets by in vitro fertilization (IVF) because of an inappropriate in vitro culture (IVC) system in pig. Therefore, the present study was conducted to investigate whether minimized exposure time in IVC can improve the pregnancy and delivery rates of piglets. Immediately after IVM, the oocytes were denuded and co-incubated with freshly ejaculated boar semen for 3.5 to 4 hours at $38.5^{\circ}C$ under 5% $CO_2$ in air. To avoid long-term exposure to in vitro state, we emitted IVC step after IVF. After that the presumptive zygotes were transferred into both oviducts of the surrogate on the same day or 1 day after the onset of estrus. Pregnancy was diagnosed on day 28 after ET and then was checked regularly every month by ultrasound examination. The 3 out of 4 surrogates were determined as pregnant (75%) and a total of 5 piglets (2 females and 3 males) were delivered at $118.3{\pm}2.5$ days of pregnancy period. In conclusion, a short-term exposure time may be an important factor in the production of IVP-derived piglets. It can be apply to the in vitro production system of transgenic pig by IVF, cloning, and pronuclear microinjection methods.

• Reproductive and Developmental Biology
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• v.38 no.2
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• pp.85-91
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• 2014

Despite many researches related with in-vitro culture of porcine spematogonial stem cells (SSCs), adherent culture system widely used has shown a limitation in the maintenance of porcine SSC self-renewal. Therefore, in order to overcome this obstacle, suspension culture, which is known to have numerous advantage over adherent culture, was applied to the culture of porcine SSCs. Porcine SSCs retrieved from neonatal testes were suspension-cultured for 5 days or 20 days, and characteristics of suspension-cultured porcine SSCs including proliferation, alkaline phosphatase (AP) activity, and self-renewal-specific gene expression were investigated and compared with those of adherent-cultured porcine SSCs. As the results, the suspension-cultured porcine SSCs showed entirely non-proliferative and significantly higher rate of AP-positive cells and expression of self-renewal-specific genes than the adherent-cultured porcine SSCs. In addition, long-term culture of porcine SSCs in suspension condition induced significant decrease in the yield of AP staining-positive cells on post-day 10 of culture. These results showed that suspension culture was inappropriate to culture porcine SSCs, because the culture of porcine SSCs in suspension condition didn't stimulate proliferation and maintain AP activity of porcine SSCs, regardless of culture periods.

• Korean Journal of Plant Resources
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• v.24 no.5
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• pp.591-598
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• 2011

This study was conducted to establish the tissue culture system for Atractylodes plant which is most frequently used in oriental medicine. Root and auxiliary bud of Dachul cv., which is Atractylodes hybrid (A. macrocephala x A. japonica), were used as target tissues for in vitro culture. In root culture, callus induction rate was higher in the treatment of BAP combined with NAA than others, however, 2-iP was more effective for callus proliferation and root induction. Although calli were effectively induced from the root and proliferated in lower concentration of cytokinin combined with higher auxin, root tissue was inappropriate for shoot regeneration. For plant regeneration with axillary bud, BAP combined with NAA was more effective than 2-iP with NAA or IBA. Number of regenerated plant per bud was 3.8, which was highest, and stem diameters was shown as 5.0mm under the conditions of 1 mg/L BAP combined with 1 mg/L NAA. Although, plant height was tend to be higher in 2-iP than BAP, number of the regenerated plant was lower via versus. Furthermore, root proliferation of regenerated plant was more effective in higher concentration of sucrose (7%) than in lower concentration (3%). In results, auxiliary bud was an efficient target tissue for producing regenerated plant of Atractylodes under the conditions of 1 mg/L BAP combined with 1 mg/L NAA and higher concentration of sucrose was effective for root proliferation of regenerated plants.