• Reproductive and Developmental Biology
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• 제29권3호
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• pp.145-147
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• 2005

Autocrine or paracrine mediators released by the early embryo are implicated in the support of embryonic development. Their mechanisms and optimal embryo density in the medium, however, are uncertain. This study was conducted to establish the optimal embryo density and culture medium volume in mouse parthenogenetic embryo culture. In experiment 1, culture of parthenogenetirally activated oocytes at a concentration of $2{\~}4$ embryos/${\mu}L$ significantly improved development to the blastoryst stage ($72{\%}{\leq}$) compared with culture at the lower ($0.2{\~}1$e mbryos/${\mu}L,\;0\~37.5\%$) and the higher ($5{\~}6$ embryos/${\mu}L,\;30\~53\%$) concentration for 120 h when the oocytes were cultured in a 5 ${\mu}L$ drop under mineral oil In experiment 2, the embryos cultured at a concentration of $2{\~}4$ embryos/${\mu}L$ in a 10 ${\mu}L$ drop ($81.1{\%}$) showed significantly higher blastocyst rates than those in a 5 ${\mu}L$ drop ($68.5{\%}$). This study optimizes in vitro culture condition by modifying embryo density and the volume of culture medium It may give appropriate level of autocrine and/or paracrine factors to enhance viability and subsequent normal development of mouse parthenogenetic embryos in vitro.

• 한국수산과학회지
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• 제43권6호
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• pp.670-678
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• 2010

An ecological investigation was conducted in winter from November 2007 to February 2008, around Wangdol Reef in the East Sea to analyze the population structure and fecundity of Chinoecetes opilio. The sex ratio was 30.11:69.89% (female:male) in November, 62.03:37.97% in December, 37.59:62.41% in January and 14.69:85.31% in February. Regression equations indicated highly positive relationships between size and weight parameters (carapace length, width, total weight). During the sampling period ovigerous females were continuously collected. The percentage of ovigerous females was 96% in November, 90.57% in December 89.80% in January, and 88.46% in February. The average number of embryos was 64,800. The regression equation between carapace width and the number of embryos was y = 2.6805x - 0.9182 ($R^2$ = 0.7166). Embryo volume increased as embryo development proceeded. The mean size of an embryo was 0.72 mm. Embryo volume ranged from 0.42 to $0.84\;m^3$ during embryo developmental stages 1 to 3.

• 한국양식학회지
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• 제20권3호
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• pp.194-198
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• 2007

Reproductive traits of Palaemon gravieri such as embryo size, number of embryo (fecundity), incubation period, larval development mode, larval development period, larval survival and larval growth were described and compared to analyze the correlation among those traits. Embryo volume is a primary factor determining other ensuing reproductive features. Egg volume was $0.042mm^3$ in the first developmental stage. Embryo volume in P. gravieri was comparatively small which is indicative of great number of embryo (y = 3.0161x + 0.0185 $R^2$ = 0.74 positive isometric relationship) and relatively long incubation period. Larvae survived from zoea 1 to post-larvae and it took 45 days at $22^{\circ}C$. Survival rate of the larvae was rather great in the early stage and thereafter steadily decreased. Daily growth rate of larvae in P. gravieri at $22^{\circ}C$ was 0.0195 mm on average. They grew steadily as time went by. Incubation period was between 10-14 days at $22^{\circ}C$. Larval development mode was almost complete planktotrophic. PNR (point of no return) appeared to be the third day on average. Survival rate of larvae without feeding declined rapidly between 3 and 4 days. Larval development period and stage frequency were 23-30 days and 11 stages which imply prolonged larval period and high mortality. The pattern of brood production followed fast successive parturial pattern. Most ovigerous female had mature ovary when they performed parturial molt soon after hatching (larval release).

• 한국수정란이식학회지
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• 제25권4호
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• pp.259-262
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• 2010

Optimization of the preimplantation mammalian embryo culture condition was widely focused on refining medium composition under the name of chemically defined media. However, recent research revealed that the alteration of physical environment can be a crucial factor to a successful embryo development. In this study, under the same embryo density, a novel culture device named oil-free micro tube culture (MTC) system was evaluated using porcine parthenogenetic embryos. The activated oocytes were placed into the 0.2 ml thin-wall flat cap PCR tube and cultured to the blastocyst stage. As a preliminary step, embryo density and culture medium volume were optimized under a standard drop culture system. The optimal embryo density range for in vitro culture was 0.5 embryos per ${\mu}l$ in $20\;{\mu}l$ drop (20.5%) and 1.0 embryos per ${\mu}l$ in $10\;{\mu}l$ drop (20.6%). Based on these results, we compared drop culture system and 'MTC' system in terms of the developmental rate to the blastocyst stage. In $20\;{\mu}l$ medium volume, the 'MTC' system showed similar blastocyst formation rate when compared with drop culture system (20.2% versus 20.5%, respectively) while the 'MTC' system showed lower blastocyst formation rate than drop culture system in $10\;{\mu}l$ one (12.7% versus 20.0%, respectively). Therefore the $20\;{\mu}l$ MTC system may be an alternative incubation system for short-distance embryo transport without carrying the $CO_2$ incubator and this provides novel embryo culture device to clinical veterinary embryologists.

• 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.

• Journal of Plant Biotechnology
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• 제37권4호
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• pp.494-504
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• 2010

본 연구에서는 고추의 소포자 배양 시 전처리 후 배지의 교환 여부, 배양 후 새 배지의 첨가 및 2층배지 사용시 하층고체배지의 양이 배의 생산에 미치는 영향을 조사하였다. 고온 전처리 후 배양배지를 첨가하여 배양하는 것보다 사용한 전처리 배지를 새 배양 배지로 교환하여 배양 하는 것이 배의 생산에 효과적 이었으며, 고온전처리 기간은 1일이나 2일에 비해 3일이 효과적 이었다. 배양 후 새 배지의 첨가는 약전처리 시에는 효과가 없었으나 소포자 전처리 시에는 배의 유기와 발달 모두 크게 향상되었다. 새 배지의 첨가 시기는 배양 4일 후가, 첨가 횟수는 1회가 배의 생산에 가장 효과적이었다. 한편 2층배지 사용 시 첨가하는 새 배지의 양은 1.5 ml이 효과적이었으며 이보다 많은 양을 첨가하는 경우 배의 발생과 발달 모두 저하되었다. 액체배지 사용 시에 비해 2층배지 사용 시 배의 발달이 좋았다. 또 2층배지 사용 시 하층고체배지의 양이 3 ml 일 때 보다는 5 ml이나 7 ml일 때 배의 발생은 감소하였으나 배의 질이 향상되었다. 이와 같은 결과들은 고추에서 다수의 정상자엽배를 생산 할 수 있는 소포자 배양시스템을 확립하는데 중요한 기초자료가 될 것이다.

• Asian-Australasian Journal of Animal Sciences
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• 제19권2호
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• pp.171-175
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• 2006

The aim of this in vitro study was to test the effect of microinjection (Mi) of foreign gene into the rabbit egg pronucleus and epidermal growth factor (EGF) addition on the blastocyst rate, the cell number and the diameter of embryos, and to determine possible relationships between embryo cell number and embryo diameter. Blastocyst rate was significantly decreased in gene- Mi (G-Mi/E0) group (63.1%) comparing to intact ones (83.5%, $p_1$<0.05). The addition of EGF at 20ng/ml (G-Mi/E20) or 200 ng/ml (GMi/ E200) to gene-Mi embryos did not affect blastocyst rate (65.6 and 55.2% resp.). As a control for Mi, the eggs were microinjected with the same volume of phosphate-buffered solution (PBS-Mi) instead of the gene construct solution. Cell numbers and embryo diameters were measured from embryo images obtained on confocal laser scanning microscope. Bonferroni-modified LSD test showed that the embryo cell number in PBS-Mi group was significantly lower ($p_1$<0.05) and in gene-Mi group was tended to decrease compared with intact embryos. Embryo diameter was not different among experimental groups. No effect of EGF given at any doses both on the cell number and embryo diameter was found. A positive correlation between cell number and embryo diameter was observed in all groups of embryos. Since embryo diameter was not changed under the influence of Mi or EGF addition in this study, this seems to be more conservative characteristics of the embryo morphology. These results suggest that the pronuclear microinjection compromises developmental potential of embryos, decreasing blastocyst rate and embryo cell number, whilst embryo diameter is not affected. No effects of EGF on studied parameters were confirmed. Declined quality of Mi-derived embryos is caused by the microinjection procedure itself, rather than by the gene construct used.

• 한국수정란이식학회지
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• 제30권3호
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• pp.171-174
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• 2015

Four pluriparous Korean black goat does were superovulated with FSH and mated with fertile bucks. Anesthetized animals were placed in lateral recumbency, then size 8 Foley catheter was inserted into the uterus through the cervix under the vaginal speculum and the balloon was inflated to fix the catheter in the uterine body. The opposite end of the catheter was connected to a 3-way and a flushing medium was infused into the uterus. Modified Dubecco's PBS with 1% FBS was used as the flushing medium. Four goats were allocated in two groups depending on the type of medium infusion into uterus. Injection group; the flushing medium was injected into uterus and the infused medium was collected by to-and-fro method using a syringe. Gravity-flow group; the flushing medium was allowed to enter the uterus by gravity flow by lifting the medium bottle and drained out of the uterus into a collecting tube. All four goats had catheter inserted through the cervix and uteri flushed successfully. The volume (recovery rate) of recovered medium varied considerably from 87 ml/200 ml (43.5%) to 148 ml/160 ml (92.5%). Nine embryos/ova in total were recovered from Gravity-flow group goats. Although the embryo recovery rate was low, the possibility of a transcervical embryo recovery in Korean black goat had been proven in this preliminary experiment.

• 전자공학회논문지CI
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• 제48권2호
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• pp.16-27
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• 2011

배아란 동물이나 식물과 같은 다세포 생물의 초기 단계를 의미한다. 배아의 단계에서 다세포 생물의 기초적인 체제가 결정되기 때문에 배아는 개체발생의 기구를 연구하는 중요한 연구대상이 된다. 생물학자들은 배아 연구를 위해 대용량의 배아 이미지 데이터를 소유하고 있으며, 이러한 대용량 데이터 중 원하는 이미지를 효율적으로 검색하기 위해서는 데이터 구조화가 필요하다. 데이터베이스 구조화를 위해 주로 사용되는 방법으로 계층적 클러스터링이 있다. 그러나 기존의 계층적 클러스터링 방법은 데이터베이스를 트리 형태로 구조화 하는 과정에서 클러스터의 크기와 클러스터 내의 객체 수를 동시에 고려하지 못하기 때문에 결과 클러스터링 트리가 경사 트리일 가능성이 매우 높다. 경사 트리인 경우 사용자가 원하는 이미지를 검색하기 위해 트리를 순회할 때 많은 시간이 걸린다. 따라서 본 논문에서는 대용량의 배아 이미지 데이터를 경사 되지 않으며 균형 상태에 가까운 트리 형태로 구조화하기 위한 방안을 제시한다. 제안하는 방안은 데이터베이스 내에 저장된 배아 이미지를 그래프로 변환하고 반복적으로 그래프 분할 알고리즘을 적용하여 클러스터를 생성한다. 이 때 클러스터의 크기와 클러스터 내의 객체 수를 동시에 고려하여 특정 클러스터의 크기가 지나치게 커지거나 객체 수가 많아지는 것을 방지한다. 실험을 통해서 제안하는 방안의 우수성을 규명하고 시각화 툴을 제공하여 사용자가 원하는 배아 이미지를 쉽게 찾을 수 있도록 돕는다.

• 한국수정란이식학회:학술대회논문집
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• 한국수정란이식학회 2003년도 제3회 발생공학 국제심포지움 및 학술대회
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• pp.98-98
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• 2003