• Journal of Korean Society of Forest Science
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• v.87 no.1
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• pp.57-61
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• 1998

Somatic embryo induction, plant regeneration, and field establishment were investigated from tissue cultured winter buds of a 10-year-old tree Aralia elata. Embryogenic calli were obtained from cultures of winter buds on MS medium supplemented with 2,4-D. A number of somatic embryos were regenerated from the calli on an embryo induction medium supplemented with 2,4-D and BA. Although abnormal somatic embryos were frequently observed, most of the embryos formed were morphologically normal. All somatic embryos at the later stage of maturity germinated successfully, but only 14% of them could be developed into plantlets on MS basal medium. The plants regenerated from the somatic embryos survived well in the field (survival rates : more than 95%) and have grown normally for three years after transplanting.

• Korean Journal of Plant Tissue Culture
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• v.22 no.2
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• pp.77-81
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• 1995

When cultured on MS medium supplemented with 0.5 mg/L NAA alone or 1.0 mg/L 2,4-D and 0.5 mg/L BA, immature zygotic embryos of Stewartia koreana formed embryogenic calli and somatic embryos. In investigate effect of sucrose concentration on somatic embryo development, embryogenic calli were transferred to MS basal medium containing 1.5,3, 6 or 9% sucrose. The greatest frequency of somatic embryos was obtained on medium containing 6% sucrose. However addition of 1.5 or 9% sucrose to medium inhibited somatic embryo germination and development into normal plantlet After 5 weeks of hardening culture on medium containing 6% sucrose, somatic embryos were transferred to half strangth MS medium supplemented with 0.1% charcol, wherein these embryo developed into the normal plantlets.

• Journal of Embryo Transfer
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• v.17 no.2
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• pp.109-115
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• 2002

The present study was conducted for the production of transgenic cloned cows those secrete human lactoferricin into milk by somatic cell nuclear transfer (NT). To estimate detrimental effects of gene transfection on transgenic cloned embryo production, development rates of NT embryos were compared between transfected and non-transfected cumulus and ear fibroblast cells. An expression plasmid for human lactofericin (pbeta-LFC) was constructed by inserting a bovine beta-casein promoter, a green fluorescent protein (GFP) marker gene, and human lactoferricin target gene into a pcDNA3 plasmid. Two bovine somatic cell lines (cumulus cell and ear fibroblast) were established and transfected with the expression plasmid using a liposomal transfection reagent, Fugene6 as a carrier. Cumulus cell and ear fibroblast were transfected at the passage of 2 to 4, trypsinized and GFP-expressing cells were randomly selected and used for somatic cell NT. Developmental competences (rates of fusion, cleavage, and blastocyst formation) in bovine transgenic somatic cell NT embryos reconstructed with non-transfectecd cells were significantly higher than those from transfected cells in cumulus cell and ear fibroblast (P＜0.05). This study indicated that transfection of done. cell has detrimental effect on embryo development in bovine transgenic NT.

• Korean Journal of Plant Tissue Culture
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• v.28 no.1
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• pp.21-24
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• 2001

Peucedanum japonicum $T_{HUNB}$ used as a edible and medicinal plants was investigated for in uitro regeneration. Callus formation occurred on leaf and stem explant cultures and showed spontaneous embryogenic and organogenic capability on MS basal medium supplemented with 0.1~5 mg/L NAA and 0~10 mg/L BA in dark. The regeneration was highest on the condition supplemented with 2.5 mg/L NAA and 10 mg/L BA. Development of the somatic embryo progressed through the globular, heart-shaped, torpedo-shaped and cotyledonary stage, typical of zygotic embryos. When the first somatic embryos was cultured on the medium supplemented with 0.2 mg/L NAA, secondary somatic embryo were induced with higher frequency on the hypocotyl then on the cotyledon and root.t.

• Proceedings of the Korean Society of Embryo Transfer Conference
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• 2002.11a
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• pp.75-75
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• 2002

The present study was conducted for the production of transgenic cloned cows by somatic cell nuclear transfer (SCNT) that secrete human prourokinase into milk. To establish an efficient production system for bovine transgenic SCNT embryos, the offset was examined of various conditions of donor cells including cell type, size, and passage number on the developmental competence of transgenic SCNT embryos. An expression plasmid far human prourokinase (pbeta-ProU) was constructed by inserting a bovine beta-casein promoter, a green fluorescent protein (GFP) marker gene, and a human prourokinase target gene into a pcDNA3 plasmid. Three types of bovine somatic cells including two adult cells (cumulus cells and ear fibroblasts) and fetal fibroblasts were prepared and transfected using a lipid-meidated method. In Experiment 1, developmental competence and rates of GFP expression in bovine transgenic SCNT embryos reconstructed with cumulus cells were significantly higher than those from fetal and ear fibroblasts. In Experiment 2, the effect of cellular senescence in early (2 to 4) and late (8 to 12) passages was investigated. No significant differences in the development of transgenic SCNT embryos were observed. In Experient 3, different sizes of GFP-expressing transfected cumulus cells [large (>30 ${\mu}{\textrm}{m}$) or small cell (<30 ${\mu}{\textrm}{m}$)] were used for SCNT. A significant improvement in embryo development and GFP expression was observed when small cumulus cells were used for SCNT. Taken together, these results demonstrate that (1) adult somatic cells could serve as donor cells in transgenic SCNT embryo production and cumulus cells with small size at early passage were the optimal cell type, and (2) transgenic SCNT embryos derived from adult somatic cells have embryonic development potential.

• Korean Journal of Medicinal Crop Science
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• v.7 no.1
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• pp.1-6
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• 1999

Basic informations for direct somatic embryo formation in Rehmannja glutinosa Lib. were obtained in 500ml erlenmyer flask. The ratio of ammonium to nitrate nitrogen of 825(mg/l) : 1900(mg/l) was proper condition for somatic embryo formation from stem and petiole explants and 3% sucrose was the most effective carbon source. Full strength MS medium with 2mg/l BA was better than LS medium for somatic embryogenesis. The initial pH 5.7 of medium(full strength MS with 2.0mg/l BA and 0.1mg/l NAA) was good for embryo production. Potassium ion was taken up rapidly within 2 weeks. while $Ca^{++}$ and $Mg^{++}$ ion contents were almost constant during culture period. Sucrose hydrolysis occurred throughout the culture, while glucose and fructose were absorbed simultaneously from the third week of culture.

• Plant Biotechnology Reports
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• v.2 no.1
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• pp.87-92
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• 2008

An improved protocol for high frequency plant regeneration via somatic embryogenesis from zygotic embryo-derived cell suspension cultures of watershield (Brasenia schreberi) was developed. Zygotic embryos formed pale-yellow globular structures and white friable callus at a frequency of 80% when cultured on halfstrength MS medium supplemented with $0.3mg\;l^{-1}$ 2,4-D. However, the frequency of formation of pale-yellow globular structures and white friable callus decreased slightly with increasing concentrations of 2,4-D up to $3mg\;l^{-1}$, where the frequency reached ~50% of the control. Cell suspension cultures from zygotic embryoderived white friable callus were established using half-strength MS medium supplemented with $0.3mg\;l^{-1}$ 2,4-D. Upon plating of cell aggregates on half-strength MS basal medium, approximately 8.3% gave rise to somatic embryos and developed into plantlets. However, the frequency of plantlet development from cell aggregates was sharply increased (by up to 55%) when activated charcoal and zeatin were applied. Regenerated plantlets were successfully transplanted to potting soil and grown to normal plants in a growth chamber. The distinctive feature of this study is the establishment of a high frequency plant regeneration system via somatic embryogenesis from zygotic embryo-derived cell suspension cultures of water-shield, which has not been previously reported. The protocol for plant regeneration of watershield through somatic embryogenesis could be useful for the mass propagation and transformation of selected elite lines.

• Journal of Embryo Transfer
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• v.26 no.1
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• pp.85-89
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• 2011

Somatic cell nuclear transfer (SCNT) is a useful tool for reproducing genetically identical animals or producing transgenic animals. Many reports have demonstrated that the efficiency of animal cloning by SCNT requires reprogramming of the somatic nucleus to a totipotent like-state. The SCNT-related reprogramming might mimic the natural reprogramming process that occurs during normal mammalian development. However, recent evidence indicates that the reprogramming event by SCNT is incomplete. In this study, the traditional SCNT procedure (TNT) was modified by injecting donor nuclei into recipient cytoplasm prior to the enucleation process to expose the donor nucleus before removing the karyoplast containing the chromosomes of the oocytes which might possess additional reprogramming factors, and this modified technique was named as reversing the usual order of SCNT (RONT). Other procedures including activation and in vitro culture were the same as TNT. Contrary to expectations, the rate of blastocyst development was not different significantly between RONT and TNT (8.6% and 7.9%, respectively). However, duration of micromanipulation performed by the same technician and equipments was remarkably reduced because the ruptured oocytes after nuclear injection were excluded from the enucleation process. This study suggests that RONT, a simplified SCNT protocol, shortens the duration of SCNT procedure and this less time-costing protocol may enable the researchers to perform murine SCNT easier.

• Proceedings of the Korean Society of Embryo Transfer Conference
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• 2002.11a
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• pp.43-55
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• 2002

Cloning by nuclear transfer in mammals using somatic cells has enormous potential applications. However, somatic cloning has been inefficient in all species in which NT is successful. High abortion and fetal death rates have been observed. These developmental defects have been attributed to incomplete nuclear reprogramming by the somatic cloning process. In this review, we will discuss studies conducted in our labs to understand the nuclear reprogramming process.

• Proceedings of the Plant Resources Society of Korea Conference
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• 2021.04a
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• pp.51-51
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• 2021

This study was conducted to develop a somatic embryogenesis protocol for the Cnidium officinale Makino difficult to seed propagation. The immature flowers were used as explants. The concentration of a 2,4-D 1.0mg/L was found to be optimal concentration for induction of embryogenic callus and somatic embryos. Addition of 0.3mg/L, 0.5mg/L and 1.0mg/L to the embryo germination medium promoted somatic embryo germination. Among four concentrations, GA₃ 1.0mg/L were superior to others. Shoots were transferred to hormone-free MS medium after 2 months of culture in the dark. We obtained an optimized protocol for the regeneration of C. officinale.