• Plant Biotechnology Reports
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• v.2 no.1
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• pp.33-39
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• 2008

In this paper, we would like to show unexpected morphogenic potential of cell suspensions derived from seedling explants of Gentiana kurroo (Royle). Suspension cultures were established with the use of embryogenic callus derived from seedling explants (root, hypocotyl and cotyledons). Proembryogenic mass proliferated in liquid MS medium supplemented with $0.5mg\;l^{-1}$ 2,4-D and $1.0mg\;l^{-1}$ Kin. The highest growth coefficient was achieved for root derived cell suspensions. The microscopic analysis showed differences in aggregate structure depending on their size. To assess the embryogenic capability of the particular culture, 100 mg of cell aggregates was implanted on MS agar medium supplemented with Kin ($0.0-2.0mg\;l^{-1}$), $GA_3$ ($0.0-2.0mg\;l^{-1}$) and AS ($80.0mg\;l^{-1}$). The highest number of somatic embryos was obtained for cotyledon-derived cell suspension on $GA_3$-free medium, but the best morphological quality of embryos was observed in the presence of $0.5-1.0mg\;l^{-1}$ Kin, $0.5mg\;l^{-1}$ $GA_3$ and $80.0mg\;l^{-1}$ AS. The morphogenic competence of cultures also depended on the size of the aggregate fraction and was lower when size of aggregates decreased. Flow cytometry analysis reveled luck of uniformity of regenerants derived from hypocotyl suspension and 100% of uniformity for cotyledon suspension.

• Proceedings of the KSAR Conference
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• 2002.06a
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• pp.51-51
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• 2002

Despite of importance of integrated events of nucleus and microtubule remodeling in nuclear transferred embryos with somatic cells, little information is available on this subject. In this study we compared chromatin, r-tubulin and microtubule organization in porcine oocytes following somatic cell nuclear transfer and parthenogenetically activation in order to clarify nuclear remodeling process and to demonstrate centrosome inheritance during nuclear transfer. (omitted)

• Proceedings of the Korean Society of Embryo Transfer Conference
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• 2002.06a
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• pp.19-25
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• 2002

Researches on manipulation pluripotent stem cells derived from blastocysts or promordial germ cells (PGCs) have a great advantages for developing innovative technologies in various fields of life science including medicine, pharmaceutics, and biotechnology. Since the first isolation in the mouse embryos, stem cells or stem cell-like colonies have been continuously established in the mouse of different strains, cattle, pig, rabbit, and human. In the animal species, stem cell biology is important for developing transgenic technology including disease model animal and bioreactor production. ES cell can be isolated from the inner cell mass of blastocysts by either mechanical operation or immunosurgery. So, mass production of blastocyst is a prerequisite factor for successful undertaking ES cell manipulation. In the case of animal ES cell research, various protocol of gamete biotechnology can be applied for improving the efficiency of stem cell research. Somatic cell nuclear transfer technique can be applied to researches on animal ES cells, since it is powerful tool for producing clone embryos containing genes of interest. In this presentation, a brief review was made for explaining how somatic cell nuclear transfer technology could contribute to improving stem cell manipulation technology.

• Korean Journal of Plant Tissue Culture
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• v.21 no.3
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• pp.177-182
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• 1994

Cotyledon segments of korean ginseng produced somatic embryos when cultured on MS basal medium, whereas plumule or excised axis explants did not. histological examination revealed that the cells in proximal region of cotyledon turned meristematic and densely cytoplasmic was composed of smaller and more densely cytiplasmic cells than the subepidermal cells. however, in the case both epidermis and subepidermal cells were almost the same in size and cytoplasmic density, the embryo originated from multiple cells.

• Journal of Plant Biology
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• v.37 no.3
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• pp.381-385
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• 1994

The highest degree of callus formation was obtained from the shoot tips of Dianthus superbus when cultured on the MS medium supplemented with 2.0 mg/L NAA and 0.5 mg/L BAP. Embryogenic calluses were obtained from the seperated friable calluses on MS medium containing 2.0 mg/L 2,4-D after 7-8 wk of culture. For plant regeneration, embryogenic calluses were selected and cultured on te proliferation medium. After 3 wk, somatic embryos appeared on MSK medium (0.5 mg/L NAA, 2.0 mg/L kinetin) and N6 medium (2.0 mg/L kinetin, 0.1 mg/LNAA, 0.1 mg/L 2,4-D and 2.0 g/L casein hydrolysate). When these somatic embryos were kept under continuous illumination, shoots were successfully regenerated on the both media. The shoots were rooted on MS medium supplemented with 2.0 mg/L NAA.

• Reproductive and Developmental Biology
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• v.31 no.2
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• pp.127-131
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• 2007

Embryonic germ (EG) cells are undifferentiated stern cells isolated from cultured primordial germ cells (PGC). These cells share many characteristics with embryonic stem cells including morphology and pluripotency. Undifferentiated porcine EG cell lines demonstrating capacities of differentiation both in vitro and in vivo have been established. Since EG cells can be cultured indefinitely in an undifferentiated state, whereas somatic cells in primary culture are often unstable and have limited lifespan, EG cells may provide inexhaustible source of karyoplasts in nuclear transfer (NT). In this study the efficiencies of NT using porcine EG and fetal fibroblast cells were compared. Two different techniques were used to perform NT. With conventional NT procedure (Roslin method) involving fusion of donor cells with enucleated oocytes, the rates of development to the blastocyst stage in EG and somatic cell NT were 16.8% (59/351) and 14.5% (98/677), respectively. In piezo-driven microinjection (Honolulu method) of donor nuclei into enucleated oocytes, the rates of blastocyst formation in EG and somatic cell NT were 11.9% (15/126) and 9.4% (9/96), respectively. Regardless of NT methods used in this study, EG cell NT gave rise to comparable rate of blastocyst development to somatic cell NT. Overall, EG cells can be used as karyoplast donor in NT procedure, and embryos can be produced by EG cell NT that may be used as an alternative to conventional somatic cell NT.

• Korean Journal of Medicinal Crop Science
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• v.10 no.3
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• pp.217-221
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• 2002

Callus induction and embryogenesis were studied in three different genotypes of Acanthopanax senticosus, to develop a protocol for somatic embryogenesis and acclimatization. Young leaf, stem, node, petiole, peduncle, flower and root explants were collected from 3-year old trees of A. senticosus accessions (Korea, Russia and Japan). Callus was obtained from all cultured explants but showed the higher rate of callus formation in flower cultured. For the three A. senticosus accessions, callus was well formd on MS media containing 2mg/ l of 2,4-D and 2mg/ l of TDZ, 4mg/ l of 2,4-D and 1mg/ l of TDZ than other treatments. For three A. senticosus accessions, when callus transferred to MS medium with 2,4-D, embryogenic cell formed. For A. senticosus accessions Korea, embryogenic cells were obtained on callus induced from petiole, stem, node and root explants, and induction rate was lower than 3%. 200mg of embryogenic callus was transferred to MS free liquid medium and somatic embryos of heart stage were obtained after 45days of culture. When somatic embryo of germination stage were transferred to solid medium, most of the embryos were regenerated into plantlets on 1/4 MS medium. Normal plants with both shoots and roots were transferred to greenhouse soil and were successfully acclimatized.

• Reproductive and Developmental Biology
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• v.32 no.2
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• pp.135-140
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• 2008

Recent studies on nuclear transfer and induced pluripotent stem cells have demonstrated that differentiated somatic cells can be returned to the undifferentiated state by reversing their developmental process. These epigenetically reprogrammed somatic cells may again be differentiated into various cell types, and used for cell replacement therapies through autologous transplantation to treat many degenerative diseases. To date, however, reprogramming of somatic cells into undifferentiated cells has been extremely inefficient. Hence, reliable markers to identify the event of reprogramming would assist effective selection of reprogrammed cells. In this study, a transgene construct encoding enhanced green fluorescent protein (EGFP) under the regulation of human Oct4 promoter was developed as a reporter for the reprogramming of somatic cells. Microinjection of the transgene construct into pronuclei of fertilized mouse eggs resulted in the emission of green fluorescence, suggesting that the undifferentiated cytoplasmic environment provided by fertilized eggs induces the expression of EGFP. Next, the transgene construct was introduced into human embryonic fibroblasts, and the nuclei from these cells were transferred into enucleated porcine oocytes. Along with their in vitro development, nuclear transfer embryos emitted green fluorescence, suggesting the reprogramming of donor nuclei in nuclear transfer embryos. The results of the present study demonstrate that expression of the transgene under the regulation of human Oct4 promoter coincides with epigenetic reprogramming, and may be used as a convenient marker that non-invasively reflects reprogramming of somatic cells.

• 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.

• Korean Journal of Plant Tissue Culture
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• v.21 no.3
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• pp.167-171
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• 1994

This study was carried out to investigate the possibility of plant regeneration through somatic embryogenesis in suspension culture of Aralia elata S. Callus was induced from the explants of leaf and petiole cultured in the MS media containing 2,4-D and TDZ. More embryogenic calli were formed from petiole and with combination treatment of 24-D and TDZ. The quarter strength MS medium was effective for increasing number of somatic embryos. Mannitol supplemented to the quarter strength MS medium, reduced somatic embryo formation but inositol increased. Normal plantlets(86%) were regenerated from mature somatic embryos in MS basal medium and 50% of those survied when transplanted to the vermiculite in greenhouse.