• 한국발생생물학회지:발생과생식
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• 제18권3호
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• pp.139-143
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• 2014

Different with other fishes, the guppies (Poecilia reticulata) is ovoviviparity, which retain their fertilized eggs within the follicle throughout gestation. The synchronously growing diplotene oocytes store nutrients in droplets and yolk, before their maturation and fertilization. The lecithotrophic strategy of development entails the provisioning of embryos with resources from the maternal yolk deposit rather than from a placenta, it allows the extracorporeal culture of guppy embryo. Studies on their early development of live bearers like the guppy including lineage tracing and genetic manipulations, have been limited. Therefore, to optimize conditions of embryo in vitro culture, explanted embryos from pregnant females were incubated in embryo medium (L-15 medium, supplemented with 5, 10, 15, 20% fetal bovine serum, respectively). We investigated whether the contents of FBS in vitro culture medium impact the development of embryos, and whether they would hatch in vitro. Our study found that in 5% of FBS of the medium, although embryos developed significantly slower in vitro than in the ovary, it was impossible to exactly quantify the developmental delay in culture, due to the obvious spread in developmental stage within each batch of eggs, and embryos can only be maintained until the early-eyed. And although in culture with 20% FBS the embryos can sustain rapid development of early stage, but cannot be cultured for the entire period of their embryonic development and ultimately died. In the medium with 10% and 15% FBS, the embryos seems well developed, even some can continue to grow after follicle ruptures until it can be fed. We also observed that embryonic in these two culture conditions were significantly different in development speed, in 15% it is faster than 10%. But 10% FBS appears to be more optimizing condition than 15% one on development process of embryos and survival rate to larvae stage.

• 식물조직배양학회지
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• 제24권5호
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• pp.257-262
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• 1997

Brassica와 Raphanus 간에 속간 교잡종을 육성하기 위해서 34개의 교배 조합을 설정한 다음 수분 12일이 경과된 배주를 BA와 GA 1.0 ㎎/L가 첨가된 MS배지에 접종하고 배의 발아율을 조사하였다. BA나 GA 첨가는 배의 발아에 큰 영향을 미치지 않았다. 무 속의 '내병총태'와 '중국청피'를 부계로 교잡한 조합에서 배의 발아율이 낮았으나 갓(Brassica juncea)을 부계로 교잡한 조합에서는 비교적 배의 발아율이 높았다. 이러한 결과는 배주배양시 배지에 첨가한 casein hydrolysate, malt extract, BA, kinetin 및 glutamine이 배의 발아에 미치는 영향보다는 부계나 모계의 교잡조합에 따라 배의 발아율에 상당한 차이가 있다는 것을 의미한다.

• 한국수정란이식학회지
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• 제12권2호
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• pp.151-159
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• 1997

In vitro fertilization(IVF) derived morula and blastocyst embryos were bisected by a simple method and cultured in vitro without zona pellucida And also bisected embryos were frozen-thawed and cultured in vitro) to evaluate the survival rate. The results obtained were as follows : The average number of grade I or II immature follicular oocytes recovered by slicing method per ovary was 11.9 from 142 ovaries. Following in vitro fertilization, the rates of cleavage and in vitro development to morula and blatocyst were 61.7 and 32.2% respectively. The successful bisection rate of IVE embryos was 67.51%, and the embryos of blastocyst stage were bisected successfully at significantly(P

• 한국작물학회지
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• 제40권3호
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• pp.308-313
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• 1995

본 시험은 미숙종자의 배배양을 이용한 보리의 세대촉진을 위하여, 재배조건(15/10, 25/15$^{\circ}C$)과 등숙기간에 따른 미숙종자의 형질을 조사하고 이들 형질들간의 상호 관계를 구명함과 동시에, 미숙종자의 배배양시 유묘의 생육 반응을 검토하였던 바, 그 결과를 요약하면 다음과 같다. 1. 출수 후 21일이 경과한 미숙종자의 배장은 이삭의 하부보다는 중상부가, 저온(15/10$^{\circ}C$)보다는 고온(25/15$^{\circ}C$) 조건에서 길었는데, 특히 저온 조건의 하부에서 짧았다. 그러나 부위별 차이보다는 생육 온도 조건에 따른 차이가 컸다. 2. 배배양 후 유아장과 근수 및 근장은 고온의 경우 이삭 부위별 차이가 적었으나, 저온조건에서는 중, 상부보다 하부의 생육이 현저히 불량하였다. 3. 등숙 기간에 따른 배장의 변리는 고온 조건에서 21일경에 최장에 이르러 더이상 신장하지 않았지만 저온 조건에서는 29일까지도 계속적으로 신장하여 저온 조건의 29일배가 고온 조건의 17일배와 유사한 경향치를 보였다. 4. 고온 조건에서 17일이 경과한 배의 배장은 입폭, 유아장, 근수 및 근장과 정의 상관이 인정되었으나, 21일이 경과한 것은 어느 형질과도 상관이 인정되지 않았다. 5. 미숙종자의 배배양을 이용한 세대촉진시에는 25/15$^{\circ}C$의 고온 조건에서 출수후 17일이 경과한 배를 이용하는 것이 적당할 것으로 보였다.

• 한국가축번식학회지
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• 제15권2호
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• pp.103-107
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• 1991

This study was carried out to investigate the survival rate in vitro culture after frozen-thawed to used DMSO(dimethyl sulfoxide), glycerol and ethylene glycol of cryoprotective agents at the zona pellucida removed and encased into alien bisected embryo of the mouse early embryos. The results obtained from this study were as follows : 1. The survival rate of in vitro culture after frozen-thawed to used cryoprotective agents of three kinds at the zona pellucida removed bisected morula was 46.6%, 35.8% and 27.3%, total or mean were 36.6%, respectively. 2. The survival rate of in vitro culture after frozen-thawed to used cryoprotective agents of three kinds at the encased into alien bisected morula was 70.6%, 65.3% and 66.4%, total or mean were 67.4%, respectively. 3. The survival rate of in vitro culture after frozen-thawed to used cryoprotective agents of three kinds at the zona pellucida removed bisected blastocysts was 50.4%, 36.7% and 30.4%, total of mean were 39.2%, respectively. 4. The survival rate of in vitro culture after frozen-thawed to used cryoprotective agents of three kinds at the encased into alien bisected blastocysts was 71.1%, 66.7% and 63.9%, total or mean were 67.2%, respectively.

• 식물조직배양학회지
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• 제26권1호
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• pp.49-52
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• 1999

고추 (Capsicum annuum L.)의 일대잡종을 이용하여 모본의 유전자형 및 생태형이 약배양 효율에 미치는 영향을 조사하였다. 모본의 유전자형에 따라 약배양 효율에 차이를 보였다. 또한 약배양 효율은 모본으로 배 발생률이 낮은 계통보다 높은 계통을 사용한 교배조합에서 높아 약배양 효율에 세포질 효과가 있는 것으로 나타났다. 또한 생태형에 따른 배 발생 효율은 재래종$\times$피망 교배조합에서 배 발생률이 17.8~46.1%로 가장 높았고, 피망$\times$재래종 조합은 5.4~8.5%의 배발생률을 보여 가장 낮았다. 재래종 $\times$남방계, 남방계 $\times$재래종, 남방계 $\times$피망, 피망 $\times$남방계 교배조합의 배 발생률은 10.5～23.1%로 비교적 중간값을 보였다.

• 한국수정란이식학회지
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• 제24권2호
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• pp.105-108
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• 2009

This study was designed to investigate the effect of kinetin on in vitro development of parthenogenetic porcine oocytes exposed to demecolcine prior to activation. In vitro matured metaphase II stage oocytes were incubated in 0 or 2 ${\mu}$g/ml demecolcine supplemented defined culture medium for 3 h and the oocytes were activated electrically. The parthenogenetic porcine embryos were then cultured in 0 or 200 ${\mu}$M kinetin supplemented defined culture medium for 7 days. Regardless of demecolcine treatment, kinetin supplementation increased blastocyst rates significantly (7.0% versus 12.1% and 4.9% versus 8.5%; Control versus Kinetin and Demecolcine versus Kinetin + Demecolcine, respectively, p<0.05). Demecolcine treatment before activation tended to decrease blastocyst rates regardless of kinetin supplementation although it is not statistically significant. Total cell numbers in the blastocysts also tended to be elevated in embryos when supplemented with kinetin, however only the result between Kinetin and Demecolcine groups is statistically significant (37.6 ${\times}$ 7.2 versus 28.1 ${\times}$ 9.5, respectively, p<0.05). In conclusion, the present report shows that kinetin enhances developmental competence of parthenogenetic porcine embryo regardless of demecolcine pre-treatment before parthenogenetic activation when they were developed in defined culture condition.

• Journal of Forest and Environmental Science
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• 제33권3호
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• pp.197-201
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• 2017

Populus. caspica L. is an Iranian indigenous poplar species which naturally distributed in the northern part of country. Unfortunately, overuse has removed many of the stems of better form, so that natural stands now usually appear small and crook. Therefore genetic variation for selection of new superior clone of this species is needed. Conventional hybridization system is currently used to induce genetic variation in poplar species but incompatibility barriers have been observed between them. In vitro ovule embryo culture was used to overcome incompatibility obstacle for interspecific hybridization between Populus caspica L. with Populus deltoids L.62/75. Female flowers of Populus caspica L. have artificially been pollinated with pollen grain of P. deltoides 62/75 in one direction using twig and pot crossing system. Ovaries at different ages (7, 14 and 21 days after pollination) were disinfected through 70% ethanol for 1 minute, 5% of sodium-hypochlorite solution for fifteen min followed by three time rising with sterile distil-water. Isolated ovaries were then transferred to MS hormone free medium containing 30 and 60 g/L sucrose for embryo development and germination. Collected data have been analyzed by two factorial experimental designs. The results indicated that there were significant differences between age of embryos for development and germination at ${\alpha}=0.01%$. Highest embryo germination (45%) was observed from 21 days old ovaries. No significant differences were observed between MS culture media containing 30 and 60 g/L for percentages of ovary-embryo germination and number of germinated embryo per ovary at ${\alpha}=0.05%$. Fourteen percentage of embryo germination obtained in MS medium supplemented with 60 g/L sucrose, while only 35% of isolated ovaries were able to germinate in MS containing 30 g/L sucrose. Induced plantlets in 4 cm height were transferred into pots containing soilless (1:1:1 peat, per lit and vermiculite) medium for acclimatization. After successful acclimatization, plants were delivered to nursery.

• Clinical and Experimental Reproductive Medicine
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• 제49권1호
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• pp.26-32
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• 2022

Objective: We investigated the effect of supplementing fertilization medium and/or culture medium with astaxanthin (AST) on the two phases of in vitro fertilization: gamete fertilization and embryo development. Methods: Mouse cumulus-oocyte complexes were divided into four groups with 5 µM AST added to the fertilization medium (group 3, n=300), culture medium (group 2, n=300), or both media (group 4, n=290). No AST was added to the control group (group 1, n=300). Results: The fertilization rate was significantly higher (p<0.001) in the groups using AST supplemented fertilization medium (group 3, 79.0%; group 4, 81.4%) than those without AST (group 1, 56.3%; group 2, 52.3%). The blastocyst rate calculated from the two-cell stage was significantly lower (p<0.001) in the groups using AST-supplemented embryo culture medium (group 2, 58.0%; group 4, 62.3%) than in those without AST (group 1, 82.8%; group 3, 79.8%). The blastocyst rate calculated from the number of inseminated oocytes was highest in group 3 (189/300, 63.0%) and lowest in group 2 (91/300, 30.3%) with statistical significance compared to other groups (p<0.001). There were significantly higher numbers of cells in the inner cell mass and trophectoderm, as well as significantly higher total blastocyst cell counts, in group 3 than in the control group. Conclusion: An increased blastocyst formation rate and high-quality blastocysts were found only in the fertilization medium that had been supplemented with AST. In contrast, AST supplementation of the embryo culture medium was found to impair embryo development.

• Journal of Animal Science and Technology
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• 제59권11호
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• pp.24.1-24.14
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• 2017

Over the past decades, in vitro culture media have been developed to successfully support IVF embryo growth in a variety of species. Advanced reproductive technologies, such as somatic cell nuclear transfer (SCNT), challenge us with a new type of embryo, with special nutritional requirements and altered physiology under in vitro conditions. Numerous studies have successfully reconstructed cloned embryos of domestic animals for biomedical research and livestock production. However, studies evaluating suitable culture conditions for SCNT embryos in wildlife species are scarce (for both intra- and interspecies SCNT). Most of the existing studies derive from previous IVF work done in conventional domestic species. Extrapolation to non-domestic species presents significant challenges since we lack information on reproductive processes and embryo development in most wildlife species. Given the challenges in adapting culture media and conditions from IVF to SCNT embryos, developmental competence of SCNT embryos remains low. This review summarizes research efforts to tailor culture media to SCNT embryos and explore the different outcomes in diverse species. It will also consider how these culture media protocols have been extrapolated to wildlife species, most particularly using SCNT as a cutting-edge technical resource to assist in the preservation of endangered species.