• 한국자원식물학회지
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• 제17권1호
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• pp.7-13
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• 2004

인삼 천풍의 배(embryo)로부터 부정근을 유도하고 배양배지조성, 식물 호르몬과 인삼염의 농도가 인삼부정근의 생장과 ginsenosides의 생성에 미치는 영향을 조사하였다. 배지 종류별 인삼 부정근의 생장량은 B5, WPM, MS 배지 순으로 높았으며, ginsenosides 함량은 B5배지에서 가장 높았으며 panaxa triol계 사포닌 보다 panaxa diol계 사포닌이 증가하였다. 식물 생장조절제 처리구에서의 인삼부정근의 생장량은 kinetin 0.1 mg/L에서 높게 나타났으며 , ginsenosides 함량은 kinetin 0.01 mg/L에서 오히려 높게 나타났다. 특히 kinetin 0.01 mg/L 첨가에서 ginsenoside Rb$_1$의 함량이 크게 증가하였다. KH$_2$PO$_4$를 추가 첨가 할때에는 부정근의 생장에 2.5 mM에서 대조구에 비해 양호하였으나, ginsenosides의 함량은 1.25 mM에서 높았으며 , 특히 ginsenosides Re의 급격한 증가로 인하여 panaxatriol계 사포닌의 함량이 증가되었다.

• 한국동물번식학회:학술대회논문집
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• 한국동물번식학회 2002년도 춘계학술발표대회 발표논문초록집
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• pp.44-44
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• 2002

본 연구는 돼지의 미성숙 난포란을 체외에서 성숙, 수정시킨 후, 생산된 체외 수정란을 NCSU 23 체외배양액에 항산화제(NAC, ebselen 및 glutathione) 와 성장 인자 (EGF, PDGF)의 첨가가 돼지 체외 수정란의 체외 발육에 미치는 영향을 검토하였다. NAC을 0, 0.5, 1.0 및 2.0 mL 첨가하여 체외배양한 결과, 상실배기 이상 발육된 체외발육성적이 1.0mL과 2.0 mM 첨가구가 여타구보다 유의하게 높은 체외발육성적을 나타냈다.(p〈0.05). (중략)

• 생약학회지
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• 제13권3호
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• pp.116-121
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• 1982

In order to investigate the side-effects of crude drugs, twenty drugs have been tested for the teratogenic effect in rats. Among seven drugs contained alkaloid as their ingredients, no one showed teratogenic effect, but Veratri rhizoma showed embryo-toxic as revealed by severe retardation in growth of the fetuses. The other thirteen drugs which have been used freguently in oriental medicines exhibited no teratogenic effect. Cyclophosphamide used as a reference compound showed severe malformation and retardation in the growth of rat fetuses. These findings suggest that the drug extracts adopted for the study might have no teratogenic effect in the rats.

• 한국자원식물학회지
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• 제16권1호
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• pp.74-81
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• 2003

인삼 종자로부터 적출한 embryo로부터 multi-shoot를 얻기 위하여 CPA와 BA를 단독 및 복합처리 하여 각각의 유기 양상을 알아본 결과 CPA 0.5mg/ t 에 BA 1.0 mg/ t 복합처리가 multi-shoot를 얻는데 가장 효과적인 것으로 나타났으며, 식물호르몬 종류를 달리한 처리구에서는 특히, 2,4-D 1mg/ t 와 kinetin 0.5mg/ t 처리구에서 한 개의 embryo 당 4∼6개의 shoot를 유기시킬 수 있었다. 또한 인삼의 형질전환을 위해 인삼 자엽으로부터 pre-embryoid를 유기하기 위한 가장 최적의 조건은 2,4-D 1mg/ t 과 kinetin 0.5mg/ t 복합처리구에서 가장 효과적이었다. 기내배양에 의해 형성된 shoot로부터 잎의 포함여부에 따른 petiole, 그리고 embryo에서 kanamycin 내성정도를 조사한 결과 잎을 포함한 petiole과 embryo는 kanamycin 100$\mu\textrm{g}$/ m1 농도에서도 다소간 생존하는 것으로 나타나 매우 높은 농도의 항생제를 사용해야 할 것으로 판단되었으며, 잎을 포함하지 않은 petiole의 적정 농도는 100$\mu\textrm{g}$/ m1 정도인 것으로 판단되었다 또한 배배양에 의한 embryo로부터 유기된 자엽 을 2,4-D 1mg/ t 과 kinetin 0.5mg/ t 복합 처리구에서 선발할 경우와 인삼 자엽으로 부터 직접 somatic embryo를 유기하여 인삼의 형질전환을 유도하기 위해서 가장 적합한 kanamycin 농도로는 75∼100$\mu\textrm{g}$/ m1이 가장 효과적인 것으로 나타났다.

• Journal of Plant Biotechnology
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• 제29권3호
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• pp.193-198
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• 2002

작약의 약 및 소포자 배양 효율을 향상시키기 위하여 배지 내에 첨가되는 PAA의 농도별 배형성 정도와 배의 형태적 변이 등에 대한 실험을 수행하여 얻어진 결과를 요약하면 다음 과 같다. 작약의 약배양에서 배지 내에 첨가되는 PAA의 농도에 따라 소포자 유래의 배형성률은 다르게 나타났는데 2 mg/L의 PAA가 첨가된 배지에서 배형성률이 가장 높게 나타났으며, PAA의 농도가 증가됨에 따라 배형성를은 낮아진 반면 캘러스 형성률은 증가하는 경향을 나타내었다. 소포자에서 유래된 배의 형태는 PAA의 농도에 따라 큰 변이를 나타내었는데 2개의 자엽을 가진 정상배의 출현빈도는 2 mg/L의 PAA가 첨가된 배지에서 가장 높게 나타났으며 PAA의 농도가 그 이상으로 증가하면 오히려 비정상배의 출현빈도가 높아지는 경향이었다. 작약의 소포자 배양에서도 2 mg/L의 PAA 첨가 배지에서 10일간 전배양된 약을 PAA (2 mg/L)를 함유한 MS액체 배지에 배양했을 때 누출된 소포자 (shed microspore)의 수도 많았고 분열률도 높았으며, 이들 소포자로부터 형성된 배의 수도 가장 많았다.

• 한국자원식물학회지
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• 제16권2호
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• pp.160-167
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• 2003

유색칼라 미숙배 배양에 의한 급속 증식체계를 확립 하고자 Southern Light 품종을 자가수분 시켜 배발육단계 및 식물생장조절제 종류와 농도가 다량의식물체 재분화에 미치는 효과를 조사하였다. 배 발육단계 별로 MS 기본배지 에 배양한 결과 globular 단계 미숙배는 반응이 없었고 몇 개의 어뢰형 배 및 초기 자업기배는 팽대 및 발아되었는데 어린 자엽기 미숙배가 87.5% 발아율을 보여 가장 효과적이었다. 한편 어린 자엽기 미숙배를 2,4-D, NAA및 BA를 단용 또는 혼용처리한 결과 2,4-D 단용 및 BA와 혼용처리배지에서는 유백색의 점액성 캘러스만이 8개월까지 증식되었고 NAA와 BA혼용처리 배지에서는 담황색의 단단한 배발생적 캘러스가 증식되어 모두 식물체로 재분화되었는데 특히 0.5 mg/L NAA와 1.0 mg/L BA혼용처리 하였을때 한개 자엽기 미숙배로 부터 25-30개의 식물체가 재분화 되었으며 동일배지에 10% coconut water를 첨가하므로 기내 식물의 급속생장이 효과적이었다. 배유래 재분화된 식물체는 vermiculite, perlite 및 모래를 1:1:1로 혼합한 토양에 이식하였으며 100% 정상식물체로 자라고 있다.

• Reproductive and Developmental Biology
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• 제36권1호
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• pp.33-37
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• 2012

Differential capacity of the parthenogenetic embryonic stem cells (PESCs) is still under controversy and the mechanisms of its neural induction are yet poorly understood. Here we demonstrated neural lineage induction of PESCs by addition of insulin-like growth factor-2 (Igf2), which is an important factor for embryo organ development and a paternally expressed imprinting gene. Murine PESCs were aggregated to embryoid bodies (EBs) by suspension culture under the leukemia inhibitory factor-free condition for 4 days. To test the effect of exogenous Igf2, 30 ng/ml of Igf2 was supplemented to EBs induction medium. Then neural induction was carried out with serum-free medium containing insulin, transferrin, selenium, and fibronectin complex (ITSFn) for 12 days. Normal murine embryonic stem cells derived from fertilized embryos (ESCs) were used as the control group. Neural potential of differentiated PESCs and ESCs were analyzed by immunofluorescent labeling and real-time PCR assay (Nestin, neural progenitor marker; Tuj1, neuronal cell marker; GFAP, glial cell marker). The differentiated cells from both ESC and PESC showed heterogeneous population of Nestin, Tuj1, and GFAP positive cells. In terms of the level of gene expression, PESC showed 4 times higher level of GFAP expression than ESCs. After exposure to Igf2, the expression level of GFAP decreased both in derivatives of PESCs and ESCs. Interestingly, the expression level of $Tuj1$ increased only in ESCs, not in PESCs. The results show that IGF2 is a positive effector for suppressing over-expressed glial differentiation during neural induction of PESCs and for promoting neuronal differentiation of ESCs, while exogenous Igf2 could not accelerate the neuronal differentiation of PESCs. Although exogenous Igf2 promotes neuronal differentiation of normal ESCs, expression of endogenous $Igf2$ may be critical for initiating neuronal differentiation of pluripotent stem cells. The findings may contribute to understanding of the relationship between imprinting mechanism and neural differentiation and its application to neural tissue repair in the future.

• Clinical and Experimental Reproductive Medicine
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• 제24권3호
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• pp.377-383
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• 1997

The effects of embryo number and incubation volume on the development of mouse embryos were evaluated. The growth rate of two-cell mouse embryos to attached blastocyst stage and the growth rate of blastocysts to early somite stage were assessed after culture in different incubation volumes and embryo densities. Embryos were collected from ICR female mice superovulated with pregnant mare serum gonadotropin and human chorionic gonadotropin and mated by ICR males. In experiment 1, groups of one, five, ten, twenty 2-cell embryos were cultured in 10-, 50-, 500-, 1000-${\mu}l$ drops of BWW media under mineral oil at $37^{\circ}C$ in a humidified atmosphere of 5% $CO_{2}$ and 95% air. As the incubation volume decreased, significantly (p<0.05) higher rates of embryos reached morular and blastocyst stage on day 3 and 4 culture, respectively. In experiment 2, groups of one, five, ten, twenty blastocysts were cultured in 1- and 2-ml volumes of CMRL 1066 media under same condition as in experiment 1. However the reverse was the result. Decreasing the number of embryos incubated per volume from 1 to 20 significantly (p<0.05) increased the number of blastocysts reaching the late egg cylinder (LEC) and early somite (ES) stage on day 6 and 8 culture, respectively, regardless of incubation volume. Blastocysts cultured in 2ml had higher (p<0.05) development rates to LEC and ES stage on day 6 and 8 culture, respectively, than embryos cultured in 1ml. Our results suggest that the effects of embryo number and incubation volume on the development of mouse embryos are stage specific and the shifting point was between hatching and EEC stage.

• Plant Biotechnology Reports
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• 제4권2호
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• pp.125-128
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• 2010

Culture conditions were established for high frequency plant regeneration via somatic embryogenesis from cell suspension cultures of Nymphoides coreana. Zygotic embryos formed pale-yellow globular structures and calluses at a frequency of 85.6% when cultured on half-strength Murashige and Skoog (MS) medium supplemented with 0.3 $mg\;l^{-1}$ of 2,4-D. However, the frequency of pale-yellow globular structures and white callus formation decreased slightly with an increasing concentration of 2,4-D up to 10 $mg\;l^{-1}$ with the frequency rate falling to 16.7%. Cell suspension cultures were established from zygotic embryo-derived calluses using half-strength MS medium supplemented with 0.3 $mg\;l^{-1}$ of 2,4-D. Upon plating onto half-strength MS basal medium, over 92.3% of cell aggregates gave rise to numerous somatic embryos and developed into plantlets. Regenerated plantlets were successfully transplanted into potting soil and achieved full growth to an adult plant in a growth chamber. The high frequency plant regeneration system for Nymphoides coreana established in this study will be useful for genetic manipulation and cryopreservation of this species.

• 대한수의학회지
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• 제38권2호
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• pp.394-400
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• 1998

Cryopreservation of embryos by vitrification is a simple method to preserve bovine embryos for subsequent embryo transfer, but embryonic viability after vitrification has been inconsistent and low compared with conventional slow freezing. The aim of the present study is to examine the effect of serum or serum albumin in a vitrification solution and epidermal growth factor(EGF) or fibroblast growth factor(FGF) on in vitro viability of bovine blastocysts frozen by vitrification. Bovine blastocysts were produced by in vitro maturation, fertilization of follicular oocytes and culture of embryos in a synthetic oviduct fluid medium(SOFM) containing BSA and 19 essential and nonessential amino acids. Blastocysts with excellent or good morphology were selected at 7 or 8 days after culture and utilized for vitrification. In experiment 1, blastocysts were vitrified in a solution containing semi-fetal calf serum(SFCS) or BSA(5 or 10mg/ml) and then their subsequent viabilities were examined by culturing thawed embryos in a SOFM containing BSA and 19 amino acids. Effect of EGF or FGF added to a SOFM containing polyvinyl alcohol(PVA) on the viability of vitrified-thawed blastocysts was investigated in experiment 2. BSA added at 5 or 10mg/ml to a vitrification solution showed significantly higher(p < 0.05) developmental rate to expanded and hatching blastocysts than SFCS, but there was no significant difference in the developmental rate to hatched blastocysts after thawing. Supplementation of a culture medium with EGF and/or FGF significantly increased(p < 0.05) embryo development to expanded blastocysts compared with control but showed no beneficial effect on the development to hatching or hatched blastocysts. Coculture of thawed embryos with granulosa cells in a TCM 199 containing 10% fetal calf serum(FCS) showed the highest developmental rate to expanded, hatching and hatched blastocysts among the groups tested. In conclusion, supplementation of a vitrification solution with BSA at 5mg/ml and culture of thawed blastocysts in a medium containing EGF and/or FGF can improve in vitro viability of bovine blastocysts frozen by vitrification.