• 한국동물번식학회:학술대회논문집
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• 한국동물번식학회 2001년도 춘계학술발표대회
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• pp.13-13
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• 2001

This study was undertaken to evaluate the effects of catalase using xanthine (X) - xanthine oxidase (XO) system on in vitro maturation and fertilization in pig. When follicular oocytes were cultured in maturation medium with X and/or XO, the maturation rates were not significantly different between in medium with and without catalase despite of different culture periods. However, significantly (P<0.05) higher maturation rates were obrained in culture with X-XO system. The rates of degenerated oocytes were increased with culture periods prolonged, and were significantly (P<0.05) higher in medium without than with catalase at 120 h of culture. On the other hand, the parthenogenetic oocytes were observed with high proportions at 72 h of culture, hut were not different in medium with and without catalase at various times of culture. In another experiment, the frozen-thawed boar spermatozoa treated with X-XO system for in vitro fertilization. The penetration rates were higher in medium with that than without catalase during the in vitro fertilization with, none (P<0.05), XO and X＋XO. On the other hand, when sperm were treated with none, X, XO and X＋XO, lipid peroxidation were higher in medium without that than with catalase. However, the changes in sperm penetration and lipid peroxidation showed opposite patterns. The sperm suspensions were also treated with X and/or XO for assay of sulfhydryl (-SH) group content. Under the above all conditions, sperm-SH group were higher detected In medium with that than without catalase. The activity of sperm binding to zona pellucida was also evaluated through binding to salt-stored porcine oocytes. In control group, sperm binding to zona pellucida were higher than in medium with X, XO and X＋XO groups. No significant differences, however, were observed between medium with and without catalase. In conclusion, the exposure of follicular oocytes and spermatozoa to X-XO system may be caused stimulating in vitro maturation and fertilization in pig. This work was supported by grant No. 2000-1-22200-001-3 from the Basic Research Program of the Korea Science & Engineering Foundation.

• 한국수정란이식학회지
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• 제14권1호
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• pp.9-15
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• 1999

This study was conducted to investigate the effect of $\alpha$-tocopherol and cysteamine with Whitten's medium in supporting the development on in vitro maturation(IVM), in vitro fertilization (IVF) and in culture(IVC) on porcine oocytes. When the immature oocytes were cultured of $\alpha$-tocopherol for 40h, the nuclear maturation rates were 39, 4, 52.5 and 54.1%, respectivley. The nuclear maturation rates of treat groups were signficantly (P<0.05) higher than those of non-treat groups. After matureation, the oocytes were inseminated in vitro in medium 199 with ejaculated spermatoza for examination of sperm penetration, polyspermy, male pronuclear(MPN) formation, and cleavage rate. Sperm penetration rates of treat higher than the control groups(P<0.05), and MPN formation rates were significantly(P<0.05) higher on treated groups (24.3~53.1%) than control groups(14.2~21.4%). After insemination, the cleavage rates at 120hr were groups higher than control groups(P<0.05).

• 한국발생생물학회:학술대회논문집
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• 한국발생생물학회 1998년도 제4차 학술발표대회 및 정기총회
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• pp.41-42
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• 1998

The changes in calcium binding protein(CBP) of mouse epididymal sperm during their post-testicular differentiation were analyzed by two-dimensional SDS-PAGE. According to dpididymal maturation, capacitation and acrosome reaction of spermatozoa, both quantitative and qualitative changes of CBPs in the epididymal sperm was detected. It suggested that the development of fertilizing ability of epididymal sperm was closely related to the changes in the CBPs profiles of sperm during epidiyaml transit.

• 한국가축번식학회지
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• 제26권1호
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• pp.17-23
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• 2002

본 연구는 지금까지 돼지 난포란 성숙을 위해서 많이 사용하고 있는 mTCM-199, mWaymouth MB 752/1 그리고 NCSU-23 성숙배지를 비교하고 액상정액을 이용한 체외수정 방법을 개발하고자 실시하였다. 미성숙 난포란은 0.5 $m\ell$의 성숙배지에 각 well 당 30~40개씩 적하하였고, 38.5$^{\circ}C$, 5% CO2, 95% 공기로 조절된 CO2 배양기에서 44시간 성숙 시켰다. 미성숙 난포란을 mTCM-199, mWaymouth MB 752/l 그리고 NCSU-23 성숙배지에서 44시간 배양한 결과 CVBD 발생율은 각각 95.6, 94.1 그리고 94.9%였으며, MH단계까지의 성숙율은 각각 92.5, 90.1 그리고 91.1%였다. 성숙배지별, GVBD 발생율과 MH까지의 성숙율간에 유의성은 인정되지 않았다. 액상정액의 제조용 정액은 90% 이상의 운동성을 가진 농후정자부분을 사용하였으며 정액 채취 후 2시간 동안에 22~24$^{\circ}C$의 실온까지 냉각시켰다.실온까지 냉각한 정액은 BTS 희석액으로 2$\times$$10^{8}$ $m\ell$ 정자농도로 조정하여 100 $m\ell$ 플라스틱병에 30 $m\ell$씩 주입하여 17$^{\circ}C$에서 5일간 보관하였다. 5일 보관 후 운동성이 70% 이상인 정자를 체외수정에 이용하였다. 성숙 후 cumulus cell들이 제거된 성숙난포란은 0.5 $m\ell$의 mTCM-199 또는mTBM 수정배지에 30~40개씩 적하하고, 최종정자농도를 2$\times$$10^{6}$$m\ell$되도록하여 6시간 동안 수정시켰다. 체외수정시킨 수정란들은 0.5 $m\ell$의 NCSU-23 배양배지에서 수정 후 6시간 배양하여 정자침입율, 다정자침입율 그리고 웅성전핵형성율을 조사하였고, 수정 후 45시간 배양하여 난할율을 조사하였다. NC-SU-23 성숙배지와 mTBM 수정배지를 이용하였을때 웅성전핵형성율이 48.0%로써 mTCM-199 성숙배지와 수정배지 또는 mWaymouth MB 752/1 성숙배지와 mTCM-199 수정배지를 이용하였을 때보다 웅성 전핵 형성율이 높았다. 2~4세포기까지의 난할율은 mTCM-199 성숙, 수정 및 배양배지에서 24.1%, mWaymouth 752/1 성숙배지, mTCM-199 수정 및 배양배지에서 43.6%, 그리고 NCSU-23 성숙배지, mTBM 수정배지 및 WCSU-23 배양배지를 이용한 것이 71.2%였다. 이상의 결과를 종합하면 BTS 희석액으로 17$^{\circ}C$에서 5일 보존한 액상정액으로 체외수정이 가능함을 입증하였고, NCSU-23 성숙배지, mTBM 수정배지 및 NCSU-223 배지가 미성숙 난포란의 성숙, 수정 및 배양에 우수한 배지임을 입증하였다.

• 한국수정란이식학회지
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• 제9권3호
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• pp.261-268
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• 1994

Immatured bovine follicular oocytes added with serum, hormones, granulosa cells and bovine oviduct epithelium cells were fertilized in vitro after in vitro maturation. In vitro maturation and early development capacity were examined and IVF-derived embryos were transferred and to recipients and effects of sperm treatment on in vitro capacitation were investigated. The rate of in vitro maturation was improved when they were co-culutred with granulosa cells in the TCM199 medium added with 10% FCS and hormones. The percentage of acrosome reaction was not differed between sperm treatments and sperm of above 25% under-went AR during 30 min preincubation with caffeine and heparin. The cleavage rate of oocytes in vitro fertilized in TCM199 medium added with 10% FCS and hormones, GC or BOEG higher than that in medium with 10% FCS and GC. But the rate was not significantly different between GC and BOEG The cleavage of rate oocytes cultured in medium containing serum, hormones and BOEG was 80.2% and more embryos were developed to Blastocyst (17.3%). The selected embryos were transferred to 9 recipients by surgical or nonsurgical method but did not result in pregnancy.

• Applied Microscopy
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• 제31권3호
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• pp.257-266
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• 2001

황소개구리의 정자변태과정과 정자형성단계 중에 글리코겐이 정자 성숙에 미치는 역할을 알아보기 위하여 관찰한 결과는 다음과 같았다. 정자변태과정은 핵과 세포질 소기관의 형태적 특징을 토대로 하여 3단계로 나눌 수 있었다. 특히, 정자형성단계 중에서 글리코겐은 정원세포부터 정모세포발생 단계까지는 관찰되지 많았지만, 정자변태단계인 초기 정자세포부터 성숙기까지는 정자세포의 핵과 첨체 및 세포질 내에 존재하고 있었고, 성숙한 정자의 중편부에도 위치하고 있었다. 이러한 사실은 글리코겐이 정자 성숙에 중요한 역할을 수행할 뿐만 아니라 정자의 파동운동에 에너지원으로서 중요한 역할을 할 것으로 사료된다.

• Clinical and Experimental Reproductive Medicine
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• 제44권2호
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• pp.79-84
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• 2017

Objective: Optimizing in vitro maturation (IVM) media to achieve better outcomes has been a matter of interest in recent years. The aim of this prospective clinical trial was to investigate the effects of different media on the IVM outcomes of immature oocytes at the germinal vesicle (GV) stage. Methods: A total of 400 immature oocytes at the GV stage with normal morphology were retrieved from 320 infertile women aged $31{\pm}4.63years$ during stimulated intracytoplasmic sperm injection (ICSI) cycles. They were divided into groups of homemade IVM medium (I, n = 100), cleavage medium (II, n = 100), blastocyst medium (III, n = 100), and Sage IVM medium (IV, n = 100) and cultured for 24 to 48 hours at $37^{\circ}C$. ICSI was performed, and the rates of fertilization and embryo formation were compared across the four groups. Results: In the 400 retrieved GV oocytes, the total maturation rates showed significant differences in groups I to IV (55%, 53%, 78%, and 68%, respectively, p<0.001). However, there were no significant differences in the fertilization, embryo formation, or arrest rates of metaphase II oocytes across these groups. In all groups, GV maturation was mostly completed after 24 hours, with fewer oocytes requiring 48 hours to mature (p<0.01). Moreover, the rate of high-quality embryos was higher in group IV than in the other groups (p=0.01). Conclusion: The quality of the IVM medium was found to affect clinical IVM outcomes. Additionally, blastocyst medium may be a good choice in IVM/ICSI cycles as an alternative IVM medium.

• 한국수정란이식학회지
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• 제17권3호
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• pp.239-249
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• 2002

Embryos derived from pig oocytes matured in mSOF are able to develop to blastocysts after IVF. Experiment 1 evaluated the effects of two maturation media (TCM-199 vs mSOF) on maturation rate, fertilization parameters, including penetration, polyspermy, male pronuclear formation, and the mean number of sperm penetrated per oocyte. Experiment 2 and Experiments 3 examined the effects of two maturation media on zona pellucida solubility and cortical granule distribution by transmissible electron microscopy, respectively. Experiment 4 assessed the effects of two maturation media on the in vitro embryo cleavage rate and development to blastocyst. Lastly, experiment 5 examined the cell number of blastocyst. An effect of media (P<0.05) was detected for mSOF on the mean number of sperm per oocyte. In TCM group, zona digestion time (196.5$\pm$15.5 vs 131.6$\pm$20.1 before IVF, 397.5$\pm$30.3s vs 185.3$\pm$16.4s after IVF, p<0.05) was higher in TCM-199 group. No significant effects of media was observed on cortical granule distribution between two groups by TEM. An effect (P<0.05) was observed on embryo development to blastocyst (16% vs 8%) but not on cleavage rates. No significant effects of media was observed on total cell number of blastocyst. We found that the high mean number of sperm penetrated per oocyte and the weaker zona pellucida on the basis of the digestion time was shown in pig oocytes matured in mSOF, however, porcine oocyte maturation with supplemented synthetic oviduct fluid medium (mSOF) resulted in blastocyst cell numbers comparable to those observed with Tissue Culture Medium 199.

• 한국가축번식학회지
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• 제26권1호
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• pp.1-7
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• 2002

본 연구는 mTCM-199, mWaymouth MB 752/1 그리고 NCSU-23 성숙배지에서 성숙시킨 난포란을 mTBM 수정배지에서 액상정액의 정자를 이용하여 주입정자 농도별로 체외 수정시킴으로써 액상정액을 이용한 새로운 체외수정 방법을 개발하고자 실시하였다. 미성숙 난포란은 0.5 $m\ell$의 성숙배지에 각 well 당 30~40개씩 적하하였다. 액상정액 제조용 정액은 90% 이상의 운동성을 가진 농후정자부분을 사용하였으며 정액채취 후 2시간 동안에 22~24$^{\circ}C$의 실온까지 냉각시켰다. 실온까지 냉각한 정액은 BTS 희석액으로 2$\times$$10^{8}$ $m\ell$ 정자농도로 조정하여 100$m\ell$ 플라스틱병에 30 $m\ell$씩 주입하여 17$^{\circ}C$에서 5일 보관하였다. 5일 보관후 운동성이 70% 이상인 정자를 체외수정에 이용하였다. 38.5$^{\circ}C$, 5% $CO_2$, 95% 공기로 조절된 CO2 배양기에서 44시간 성숙 후 cumulus cell들이 제거된 성숙 난포란은 0.5 $m\ell$의 mTBM 수정배지에 30~40개씩 적하하고 최종정자농도를 1, 2, 4, 6 그리고 10$\times$$10^{6}$$m\ell$되도록하여 주입하고 6시간 동안 수정시켰다. 체외수정시킨 수정란들은 수정 후 6시간 동안 0.5$m\ell$의 NCSU-23 배양배지에서 배양한 후 정자침입율, 다정자침입율 그리고 웅성전핵 형성율을 조사하였다. mTCM-199, mWaymouth MB 752/1 그리고 NCSU­23 성숙배지에서 성숙시킨 난포란을 mTBM 수정배지에서 액상정 액으로 체외수정 한 결과 NCSU-23 성숙배지에서 성숙한 난포란이 웅성전핵형성율이 높았고 다정자침입율이 낮았다. NCSU-23 성숙배지에서 성숙한 난포란을 mTBM 수정배지에서 수정할 경우 최적정자농도는 2~4$\times$$10^{6}$$m\ell$이었으며, 정자침입율은 50.8~50.9%, 다정자침입율은 13.3~19.5% 그리고 웅성전핵형 성율은 43.9~45.4%였다. 결론적으로 NCSU-23 성숙배지에서 성숙되고 mTBM 수정배지에서 수정된 난포란은 mTCM-199이나 mWaymouth MB 752/1 성숙배지에서 성숙되고 mTBM 수정배지에서 수정된 난포란보다 우수한 체외수정 결과를 나타내었다. 17$^{\circ}C$에서 5일 동안 보존한 액상정액으로 NCSU-23 배지에서 성숙한 난포란을 체외수정하기 위한 최적정자농도는 2~4$\times$$10^{6}$$m\ell$이었다.

• Animal cells and systems
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• 제11권2호
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• pp.227-234
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• 2007

Spermatogenesis, the reproductive cycle, and the size at first sexual maturity in male Mactra chinensis were investigated by cytological and histological observations. The spermatozoon exhibits a primitive type morphology and is similar to those of other bivalves in that it contains a short midpiece with four mitochondria surrounding the centrioles. The morphologies of the sperm nucleus type and the acrosome shape of this species are cylindrical and modified cap-like, respectively. The spermatozoon is approximately $40-45\;{\mu}m$ in length including the sperm nucleus (about $1.46\;{\mu}m$), acrosome (about $1.20\;{\mu}m$) and tail flagellum. The axoneme of the sperm tail flagellum consists of nine pairs of microtubules at the periphery and a pair at the center. The axoneme of the sperm tail shows a 9+2 structure. The spawning period of this species lasts from June to September, and the main spawning occurs in July and August, when the seawater temperature is greater than $20^{\circ}C$. The percentage of individual male clams at first sexual maturity was 56.5% for those whose shell lengths were 35.1-40.0 mm, and 100% for over 45.1 mm. Accordingly, harvesting clams <35.1 mm in shell length could potentially cause a drastic reduction in recruitment, and a measure indicating a prohibitory fishing size should be taken for adequate fisheries management.