• Reproductive and Developmental Biology
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• 제39권2호
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• pp.29-36
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• 2015

Pigs are considered an ideal source of human disease model due to their physiological similarities to humans. However, the low efficiency of in vitro embryo production (IVP) is still a major barrier in the production of pig offspring with gene manipulation. Despite ongoing advances in the associated technologies, the developmental capacity of IVP pig embryos is still lower than that of their in vivo counterparts, as well as IVP embryos of other species (e.g., cattle and mice). The efficiency of IVP can be influenced by many factors that affect various critical steps in the process. The previous relevant reviews have focused on the in vitro maturation system, in vitro culture conditions, in vitro fertilization medium, issues with polyspermy, the utilized technologies, etc. In this review, we concentrate on factors that have not been fully detailed in prior reviews, such as the oocyte morphology, oocyte recovery methods, denuding procedures, first polar body morphology and embryo quality.

• 한국가축번식학회지
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• 제14권1호
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• pp.84-91
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• 1990

본 실험은 돼지난포란의 체외성숙과 체외수정 효과를 높일 수 있는 방법을 찾기 위하여 시도되었으며 직경 1~2mm와 3~7mm 난포로부터 채란된 난자를 mKRB(-BSA)에 돼지발정혈청(ESS), FCS 또는 투석돼지난포액(DFF)을 첨가한 성숙배양에서 24~48시간, 37$^{\circ}C$에서 배양하였다. 성숙된 난포란은 정소상체 정자와 24시간 배양 후 전핵행성 여부를 조사하였다. 36~48시간 배양에서 50~60%의 난자가 metaphase II에 도달되었고 난포 크기(1~2mm와 3~7mm)간에 체외성숙율의 차이는 없었으나 3~7mm 난포란에서 성숙분열이 다소 빨랐다. 체외성숙배양액에 5% ESS, 15% FCS 및 DFF 첨가시 대조구보다 다소 성숙율이 높았다. 체외수정율(전핵형성)은 5% ESS와 15% FCS 첨가 성숙시킨 난포란과 체내 수정능획득 정자와의 수정에서 각각 높은 경향이 있었다. 따라서 돼지난포란의 체외성숙과 수정에 ESS, FCS 및 투석난포액이 유효한 요인이 됨을 알 수 있다.

• Clinical and Experimental Reproductive Medicine
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• 제23권1호
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• pp.1-6
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• 1996

There are a number of problems during the process of culture in vitro on fertilization and embryo development compared to those on in vivo counterparts. And the platelet activating factor (PAF), which is found not only in mammalian spermatozoa but also preembryos, is implicated on reproductive process. To improve the environment of culture on in vitro fertilization and embryo development, coculture using salpingeal epithelial cells has been considered to accept the better result on pregnancy rate. This study was designed to determine if two different culture systems, coculture alone and PAF treated coculture, are positive or negative influence on process of in vitro fertilization and embryo culture in mouse. The cell cleavage rate reached to 2-4 cell stage at 24 hours of culture is 56.81% (50/88) and 48.21%(54/112) respectively, in PAF treated group which is added PAF on coculture and in coculture group. But the rate of cells cleavage was similar in both group after 48 hours of culture. The rate of unfertilization after insemination of oocytes was higher in coculture group(55..53%) than in PAF treated group(42.37%). And in assessment of undeveped embryos, the rate of equalized cell block was similar on both, coculture alone (35.3%)and PAF treated coculture(35.5%). while unequalized cell block was higher rate in PAF treated coculture(19.4%) than coculture alone (11.8%). But the rate of cytoplasmic degeneration of undeveloped embryos was significantly higher in PAF treated coculture than coculture alone. In conclusion, we have observed that PAF treated coculture is superior in the rates of in vitro fertilization and early embryo cell cleavage compared to those in coculture alone, but there is no difference on the rates of embryo develpments, cell degeneration, cell quality in both PAF treated coculture and coculture alone when the embryo cells were continuosly cultured for 48 hours or more.

• 한국수산과학회지
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• 제42권1호
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• pp.34-40
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• 2009

In fresh water fish hatcheries and farms, Saprolegniales often cause serious mortality to the fish and their eggs. Malachite green is an effective antifungal agent, but is carcinogenic to fish and humans. Alternative antifungal agents are needed. Presently, we tested various concentrations of MBT-01108 (Opuntia ficus-indica extracts) alone and in combination with bronopol, formalin and sodium chloride (MBT-01108 mixture) on in vitro mycelial growth and in vivo remediation of adult eel, Anguilla japonica, infected with Saprolegnia sp. and fertilized eggs of chum salmon, Oncorhynchus keta, to evaluate the compounds' antifungal efficacy on eyed-egg and hatching rates. MBT-Oll08 mixtures incorporating bronopol and formalin at respective concentrations of 50 and 30 parts per million (ppm), and 100 and 20 ppm were most effective in controlling Saprolegnia in vitro and in vivo (P<0.05). Repeated daily exposures to 50 ppm and 100 ppm MBT-01108 were more effective than exposure every 2-3 days post-fertilization for the inhibition of Saprolegnia infection of rainbow trout, Oncorhynchus mykiss eggs as compared with control (0 ppm).

• Asian-Australasian Journal of Animal Sciences
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• 제15권1호
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• pp.19-25
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• 2002

The objectives of this study were to understand the possible mechanism of duck sperm toxicity induced by arsenic exposure in vivo, and to investigate the roles of the antioxidant L-ascorbic acid in ameliorating the arsenic-induced sperm impairment. To test the acute toxicity, the percentages of mortality of mature drakes treated with different concentrations of trivalent sodium arsenite, As (III), and pentavalent sodium arsenate, As (V) were measured. The LD50 value of As (III) for mature drakes was $4.89{\pm}1.49$ ppm. Although As (V) didn't cause any deaths even at a concentration of 40 ppm, the chronic toxicity of As (V) on sperm quality was shown by a decreased fertilization rate. When the concentrations of As (V) were above 0.4 ppm, fertilization rates were lower than those of 0.04 ppm and control. Drakes treated with 40 ppm of As (V) had the highest malondialdehyde (MDA) level in the testis tissue, $3.100{\pm}0.218{\mu}mole/g$ testis. This showed that 40 ppm of As (V) significantly induced lipid peroxidation in testis tissue. For the 1.2 ppm As (III) treatment, several significant effects were observed: (1) sperm motility was decreased most dramatically by $52.0{\pm}9.1$% after three days of incubation; (2) fertilization rate of artificially inseminated semen was the lowest, $26.4{\pm}15.4$; (3) the MDA concentration in testis tissue, $7.846{\pm}0.246{\mu}mole/g$ testis, was significantly higher than the others (p<0.05); (4) the sperm number, $1.17{\pm}0.40({\times}10^9)$, was significantly lower than with the 60 ppb and control treatments (p<0.05); (5) a black appearance and soft texture was observed in the testis tissue. The antioxidant L-ascorbic acid administered along with 1.2 ppm As (III) decreased the toxicity of arsenic. The ameliorating effects included: improved sperm motility, increased sperm number and fertilization rate, and decreased MDA concentration in the testis tissue. This study suggests that the toxicity of the trivalent arsenic on sperm quality is partly from free radicals generated by its metabolic pathway, and the antioxidant ascorbic acid ameliorates arsenic-caused sperm impairment.

• Clinical and Experimental Reproductive Medicine
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• 제43권2호
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• pp.119-125
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• 2016

Objective: The aim of this study was to report a case series of in vitro matured (IVM) oocyte freezing in gynecologic cancer patients undergoing radical surgery under time constraints as an option for fertility preservation (FP). Methods: Case series report. University-based in vitro fertilization center. Six gynecologic cancer patients who were scheduled to undergo radical surgery the next day were referred for FP. The patients had endometrial (n=2), ovarian (n=3), and double primary endometrial and ovarian (n=1) cancer. Ex vivo retrieval of immature oocytes from macroscopically normal ovarian tissue was followed by mature oocyte freezing after IVM or embryo freezing with intracytoplasmic sperm injection. Results: A total of 53 oocytes were retrieved from five patients, with a mean of 10.6 oocytes per patient. After IVM, a total of 36 mature oocytes were obtained, demonstrating a 67.9% maturation rate. With regard to the ovarian cancer patients, seven IVM oocytes were frozen from patient 3, who had stage IC cancer, whereas one IVM oocyte was frozen from patient 4, who had stage IV cancer despite being of a similar age. With regard to the endometrial cancer patients, 15 IVM oocytes from patient 1 were frozen. Five embryos were frozen after the fertilization of IVM oocytes from patient 6. Conclusion: Immature oocytes can be successfully retrieved ex vivo from macroscopically normal ovarian tissue before radical surgery. IVM oocyte freezing provides a possible FP option in patients with advanced-stage endometrial or ovarian cancer without the risk of cancer cell spillage or time delays.

• 한국수정란이식학회지
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• 제17권1호
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• pp.23-32
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• 2002

본 연구는 기존의 수정란 이식기술을 다각적으로 분석하고 개선하여 한우 체내수정란의 생산 및 이식기술 체계를 확립하기 위하여 수행하였으며, 결과를 요약하면 다음과 같다. 1. 배란에 미치는 FSH analogue 간의 유의성은 인정되지 않았으나 수정률, 이식 가능 수정란 및 동결수정란 생산에는 처리간에 유의성(P<0.05)이 인정되었다. 특히, Super-OV는 Foll-tropin-V 및 Embryo-S에 비해서 이식 가능 수정란의 생산율이 유의적(P<0.01)으로 낮았다. 2. 호르몬의 투여시기에 따라 공란우의 배란정도 (P<0.1), 수정률, 이식 가능 및 동결 가능 수정란의 생산(P<0.05)은 각각 유의적인 차이를 나타냈다. 특히, 발정주기 11일차의 호르몬투여는 적합하지 않은 결과를 보였다. 3. 다배란처리후 황체의 등급에 따른 수정란의 수는 좌측 또는 우측에서 6개 이상의 황체를 보인 A등급(8.06∼9.49개)의 경우가 B, C 및 D등급(2.93 ∼6.28개)보다 유의적(P<0.01)으로 높았다

• 한국동물번식학회:학술대회논문집
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• 한국동물번식학회 2001년도 발생공학 국제심포지움 및 학술대회 발표자료집
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• pp.44-46
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• 2001

In vitro produced (IVP) embryos produced by in vitro fertilization (IVF) often exhibit wide variations in developmental competence and viability, considerably more than are exhibited by embryos that develop in vivo. These anomalies in IVP embryos may be due to heterogeneity of oocyte quality, suboptimal culture conditions, disturbances in gene expression, or most likely a combination of these factors (Ho et al., 1994; Roth et al., 1994; McKiernan and Bavister, 1998; Hasler, 1998; Schramm and Bavister, 1999; Doherty et al., 2000; Hyttel et al., 2000; Niemann and Wrenzycki, 2000; Wrenzycki et al., 2001). In research studies or in clinical applications with domesticated animals, cats, non-human primates and humans, oocytes used for IVF are usually collected from a heterogeneous cohort of ovarian follicles that include oocytes which normally might not be ovulated and/or are deficient in developmental competence. (omitted)

• 한국수정란이식학회지
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• 제19권3호
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• pp.239-244
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• 2004

본 연구는 체내, 체외 및 복제를 통하여 각각 생산된 배반포를 이식하여, 수란우의 수태율, 임신기간 및 유산율과 더불어 송아지의 생시 체중과 이후 생존율을 조사하였다. 그 결과, 수태 을은 체내수정란이 56.3%로서 복제 수정란의 19.4%에 비하여 유의하게 높았으나 (p<0.05), 체외수정란의 30.0%와는 유의성이 인정되지 않았다 유산율과 임신기간은 처리군 간에 유사한 경향이었다(유산을 0, 22.2 및 16.7%; 임신기간 278.8, 289.4 및 281.4일). 한편 송아지의 체중은 복제수정란에서 유래된 송아지의 평균 39.9kg은 체내수정란에서 유래된 송아지의 평균 25.5kg에 비하여 유의하게 높았다.(p<0.05). 체내수정란이 수태된 수란우(n=9)는 모두 정상 분만하였으며, 그 송아지 는 생후 60일령까지 모두 생존하였다. 한편 체외수정란이 수태된 수란우(n=7)도 모두 정상 분만하였으나, 그 송아지 가운데 1두는 생후 48일에 사망하였다. 복제 수정란이 수태된 수란우(n=5)는 정상 분만 3두 및 제왕절개 2두를 하였다. 정상 분만된 복제송아지(n=3) 중에서 2두는 분만 직후 사망하였으나, 제왕절개로 태어난 송아지는 생후 60일까지 모두 생존하였다.

• 농업과학연구
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• 제50권4호
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• pp.941-952
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• 2023

The metabolites of agrichemicals, such as organophosphorus pesticides, are known to be more hazardous than their parent pesticides. 3,5,6-trichloro-2-pyridinol (TCP) is a major degradation product of chlorpyrifos, one of the organophosphate insecticides widely used in agriculture. In vivo or in vitro exposure to chlorpyrifos has been known to interfere with male reproductive functions, leading to reduced fertility in mammals. Therefore, this study was performed to examine the changes in the fertilization competence of boar spermatozoa exposed to TCP. Sperm samples were subjected to varying concentrations of TCP (10, 50, 100, 200 µM) and different periods of incubation. Sperm motility, motion kinematics, viability, acrosome integrity, intracellular reactive oxygen species (ROS) production, and gene expression levels (ODf2, ZPBP2, AKAP3 and AKAP4) were evaluated after exposure of the sperm to TCP. A significant dose-dependent reduction in motility was observed in sperm samples incubated with TCP compared to the controls after both incubation periods. Sperm viability was significantly decreased in samples incubated with 50, 100, and 200 µM TCP in both incubation periods. A significantly lower percentage of normal acrosomes and gene expression levels were observed in sperm samples exposed to 50, 100, and 200 µM TCP after both incubation periods, compared to the controls. There was a significant increase in the ROS production in spermatozoa incubated with 100 - 200 µM TCP after both incubation periods. Consequently, the direct exposure of boar spermatozoa to TCP interferes with sperm functions and leads to decreased fertilization. In order to identify and address the various causes of reproductive decline, the impact of chemical metabolites needs to be discussed in depth.