• Journal of Plant Biotechnology
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• 제5권1호
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• pp.43-49
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• 2003

Resistant cell lines to azetidine-2-carboxylic acid (AZCA) were selected through rice embryo culture after mutagenic treatment of callus irradiated with 30,50,70,90 and 120 Gy. The optimum AZCA concentration for the selection of resistant cell lines was 3 or 4 mM AZCA considering $LD_{50}$ and the fresh weight of callus. Survival rate of the AZCA resistant callus showed remarkable increase in the callus irradiated with 50 and 70 Gy. Regeneration rate of the AZCA resistant callus was much lower on the whole. Ninety and 120 Gy increased the regeneration rate for calli selected from 3 and 4 mM AZCA, respectively. Based on fresh weight, survival rate and regeneration for selection of the AZCA resistant cell line, 50-90 Gy was considered as the optimum range of gamma irradiation. Irradiated calli selected from AZCA were more tolerant to NaCl than those from non-irradiated calli. It suggests that elevated resistance to osmotic stress resulted from mutagenic treatment. The level of free proline content in the AZCA resistant cell line was increased up to 3.5 times compared with that in the control. Proline content in the regenerant derived from the AZCA resistant cell line also increased to 1.7 times that from the control plants regenerated from callus grown in AZCA free medium. Selection of proline overproducing cell lines by in vitro mutagenesis was successful and seems to be useful for improvement of stress tolerance in this crop.

• Journal of Plant Biotechnology
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• 제29권4호
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• pp.241-246
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• 2002

구기자나무의 접합배의 자엽절편을 MS 배지에 오옥신 (2,4-D, NAA, IAA)와 싸이토키닌 (zeatin, kinetin, BAP)를 단독 또는 조합처리하여 체세포배와 부정아를 유도하였다. 0.5 mg/L 2,4-D를 첨가한 배지에서 배발생능 캘러스를 유도되었고 이로부터 체세포배가 발생되었다. 반면에 부정아는 0.01 mg/L 2,4-D＋0.5 mg/L zeatin을 조합한 배지에서 발생하였다. 접합자배의 여러 부위별로 배양한 결과 자엽 절편이 가장 높은 형태형성을 보였으며, 기관형성은 자엽＞유근＞배축＞배 전체 순이었다. 해부학적으로 관찰한 결과 심장형배를 관찰할 수 있었고 정단분열조직과 부정아를 확인할 수 있었다.

• 한국독성학회:학술대회논문집
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• 한국독성학회 2002년도 Current Trends in Toxicological Sciences
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• pp.71-72
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• 2002

Thimerosal is a mercury-containing compound used in trace amounts to prevent bacteria and other organisms from contaminating vaccines, especially in opened multi-dose vials. The toxicity of mercury is well known and those most at risk are occurred in unborn and newborn babies.(omitted)

• 한국환경성돌연변이발암원학회:학술대회논문집
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• 한국환경성돌연변이발암원학회 2002년도 Current Trends in Toxicological Sciences
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• pp.71-72
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• 2002

• 한국독성학회:학술대회논문집
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• 한국독성학회 2002년도 Molecular and Cellular Response to Toxic Substances
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• pp.204-204
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• 2002

Alcohol drinking during pregnancy can result in abnormal fetal development including fetal alcohol syndrome (FAS). The molecular mechanisms of FAS, however, is not completely elucidated. In the present study, we evaluated the developmental toxicity of ethanol and its metabolite, acetaldehyde using post implantation whole embryo culture and determined changes of gene expression by ethanol treatment by cDNA microarray.(omitted)

• Reproductive and Developmental Biology
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• 제36권4호
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• pp.255-260
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• 2012

The purpose of this study was to assess follicular viability and competence through in vitro culture of preantral follicles isolated from vitrified mouse whole ovaries. Mouse preantral follicles were enzymatically isolated from vitrified- warmed and fresh ovaries and cultured for 10 days followed by in vitro oocyte maturation. In vitro matured oocytes were fertilized and cultured to the blastocyst stage. Five minutes pre-exposure to vitrification solution of whole ovaries had significantly higher (p<0.05) oocyte survival and maturation rates than between 10 min exposure groups. Oocyte diameter was significantly smaller (p<0.05) in the 5 and 10 min exposure groups ($69.4{\pm}2.8$ and $67.8{\pm}3.1$) when compared to that of control group ($71.7{\pm}2.1$). There was no statistical significant difference in blastocyst development rates between vitrification group (8.6%) and the fresh control group (12.0%). The mean number of cells per blastocyst was significantly lower (p<0.05) in the vitrification group ($41.9{\pm}20.2$) than in the fresh control group ($55.1{\pm}22.5$). The results show that mouse oocytes within preantral follicles isolated from the vitrified whole ovaries can achieve full maturation, normal fertilization and embryo development.

• 한국수정란이식학회지
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• 제26권2호
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• pp.123-128
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• 2011

In vitro maturation and activation of oocytes are primary steps towards biotechnological manipulation in embryology. The objectives of the present study were to determine the oocyte recovery rate per ovary, in vitro maturation rates of oocytes and rates of parthenogenetically activation of matured oocytes in Black Bengal goats. All visible follicles were aspirated to recover follicular fluid from individual ovaries (number of ovaries = 456). The immature cumulus oocyte complexes (COCs; n = 1289) were cultured in tissue culture medium (TCM)-199 supplemented with 10% (v/v) fetal bovine serum (FBS) for 27 hours at $39^{\circ}C$ with 5% $CO_2$ in humidified air. The matured oocytes (n = 248) were activated with 5 ${\mu}M$ ionomycin for 5 minutes followed by treatment with 2 mM 6-dimethylaminopurine (6-DMAP) for 4 hours. After activation, oocytes were cultured for another 14 hours in TCM-199 supplemented with bovine serum albumin (BSA) at $39^{\circ}C$ with 5% $CO_2$ in humidified air. The pronucleus formation in activated oocytes was determined by staining with 1% orcein (whole mount technique). Matured oocytes (n = 176) without activation stimuli were used as control. The mean number of oocytes recovered per ovary was $3.5{\pm}0.5$. The proportion of oocytes matured in vitro, confirmed by the presence of first polar body, was $42.1{\pm}4.7%$. Parthenogenetic activation, evidenced by formation of pronucleus, occurred in $37.2{\pm}15.8%$ of matured oocytes. No pronucleus formation was observed in control oocytes. In conclusion, a combination of ionomycin and 6-DMAP induces activation in one third of Black Bengal goats' oocytes.

• 대한수의학회지
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• 제29권3호
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• pp.343-359
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• 1989

To study the effects of ibaraki virus on preimplantation mouse embryos collected from prepubertal ICR and BALB/cByJ mice (30~40days old) by superovulation, zona pellucidaintact(ZPI) or free(ZPF) embryos(n=774) of 4- to 8-cell and morulae were exposed to $10^{5.8}$ $TCID_{50}$ of the virus up to 96 hours. The embryos were examined morphologically by observing the degeneration and hatching rates, and virologically and immunologically by determining the presence of infection with the virus, in addition, the effect of washing the embryos to remove virus possibly attached to was also investigated. The ZPI 4- to 8-cell embryos and morulae exposed to the virus showed considerably higher degeneration rate than those not exposed, for 96, and for 72 to 96 hours, respectively(p<0.01). The ZPF 4- to 8-cell embryos and morulae exposed to the virus showed considerably higher degeneration rates than those not exposed, throughout the whole culture hours in vitro (p<0.01). The ZPI 4- to 8-cell embryos and morulae not exposed to the virus showed considerably higher rates of hatched blastocyst than those exposed (p<0.01). The virus infection rates of the ZPF 4- to 8-cell embryos and morulae were significantly higher than those of the ZPI embryos according to cell culture system. The viral antigen was detected exclusively on the zona pellucida of ZPI embryos, while the antigen was evenly distributed in the blastomeres of ZPF embryos by the immunofluorescent assay. In the ZPI embryos exposed to ibaraki virus, the virus was detected in the two times-washing groups, but not in the ten times-washing groups. The results indicated that zona pellucida of murine embryos would provide an effective protection and that ten times-washing of the ZPI embryos previously exposed to the virus was effective to remove virus from the embryos.

• 한국수정란이식학회지
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• 제32권4호
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• pp.311-317
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• 2017

Cryopreservation of oocytes by vitrification technique may contribute a lot in the field of reproductive biotechnology. The objectives of the present study were to evaluate the effectiveness of two cryo-devices for vitrification of immature oocytes of indigenous zebu cows. Slaughter house derived immature cumulus-oocyte-complexes (COCs) of cows were vitrified using 15% dimethyl sulphoxide (DMSO) as cryoprotective agent (CPA) with 0.5 mol sucrose in TCM 199 supplemented with 20% FBS. Vitrification of COCs was completed after immediate plunging of COCs loaded cryotop or French mini straw into the liquid nitrogen ($LN_2$). Then the COCs containing cryotop or French mini straws were warmed in 0.25 mol sucrose and 20% FBS supplemented TCM 199 followed by in vitro culture in $50{\mu}l$ droplets of bicarbonate buffered TCM 199 supplemented with 10% FBS, pyruvate, FSH and oestradiol for 24 hrs at $39^{\circ}C$ with 5% CO2 in humidified air. After maturation culture, oocytes were denuded and examined under inverted microscope for presence of polar body as the indication of maturation. Denuded oocytes were also stained by whole mount technique using 1% orcein to examine the maturation by presence of MII chromosomes. The in vitro maturation rate was significantly (p<0.05) higher in oocytes vitrified and warmed using crytop ($47.1{\pm}6.9%$) than that of French mini straw ($15.9{\pm}12.5%$). Moreover, in vitro maturation rate was significantly (p<0.05) highe r in control oocytes (not vitrified) ($84.5{\pm}14.2%$) than that of vitrified oocytes. In conclusion, cryotop is better than French mini straw as cryo-device for vitrification of bovine immature oocytes.

• Environmental Analysis Health and Toxicology
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• 제6권1_2호
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• pp.71-82
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• 1991

포유동물의 최기형성을 연구하는데 가장 많이 사용되는 동물인 설치류중 랫트의 배자를 이용하여, egg-cylinder기를 지나 head-fold가 이루어지는 초기체절기인 9.5일경부터 뇌, 눈, 귀, 심혈관계가 형성되는 시기인 11,5일까지 2일(48시간) 동안, 체외에서 배양하였다. 그 배양의 최적조건은 실험실마다 다르기 때문에 최적조건확립을 목적으로 이 실험을 행하였다. 온도는 37$^{\circ}C$$\pm$0.2$^{\circ}C$로 유지시키고 가스는 배양초기인 9.5일~10.5일 동안을 50%$O_2$, 5% $CO_2$, 90% $N_2$가스를 공급하여 10.5일부터 6시간 동안은 20% $O_2$, 5% $CO_2$, 75% $N_2$로 하며 나머지 18시간 동안은 40% $O_2$, 5% $CO_2$, 55% $N_2$가스를 배양계에 공급했을 때 가장 좋은 결과가 나왔다. 배양배지로서는 랫트 IC 혈청을 분리하여 열비동화시켜서 사용하였다. 이종혈청이나 합성배지를 첨가해서 배양할 경우는 랫트 혈청에 비해 성장이 좋지 않은 것으로 관찰되었다. 동물실험법의 대체법으로서 배자배양법의 기본조건을 확립하여 최기형성 평가 방법으로서의 가능성을 제시하였다.