• Title/Summary/Keyword: MATURATION

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The Expression of Matrix Metalloproteinases Activated Differently on In-Vitro Maturation of oocytes Cytoplasm and Cumulus Cells in Bovine (소 난자의 체외성숙시 난구세포와 난세포질에서 다르게 발현되는 Matrix Metalloproteinases의 분석)

  • Kim, Sang-Hwan;Yoon, Jong-Taek
    • Journal of Embryo Transfer
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    • v.33 no.3
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    • pp.99-105
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    • 2018
  • To determine the differences in the in-vitro ovum maturation process of bovine, we compared the expression of MMPs in these oocytes and cumulus cell throughout oocytes maturated. In an attempt to investigate the effect of MMP activation and inhibitors in total protein of cumulus cell and, oocytes during oocytes maturation, we examined and monitored the localization and expression of MMPs (MMP-2 and MMP-9), TIMPs (TIMP-2 and TIMP-3), as well as their expression profiles (Real-time PCR, Gelatin Zymography and ELISA). Our results that the bovine oocytes MMP-2 and MMP-9 level was significantly associated with the rate of maturity of oocytes (P<0.05). In cumulus cell, MMP-2 was highly expressed in all stages of the oocyte's maturation. The final oocytes maturation exhibited strong gelatinase activity. There was no significant correlation between cumulus cell MMP-9 and the maturation rate of oocytes. However, for the oocyte cytoplasm MMP-9 expression was significant correlation to the maturation oocytes. There was no significant correlation between cumulonimbus cells MMP-9 and oocyte maturation rates; however, for oocyte cytoplasm, MMP-9 expression was significantly correlated with mature oocyte. However, the TIMP-1 and TIMP-2 protein expression patterns are not correlated with the maturation rate of the oocyte. Our results suggest that MMP different expression pattern may regulate the morphological remodeling of oocyte's in the cumulus cell. Further, the MMP-2 expression has a strong relation with a higher maturation rate of the oocyte.

Analysis of Factors related with Early Bony Maturation During the Growth Period (성장기 동안의 조기 골성숙 관련 인자들;비만도와 모발영혈검사를 중심으로)

  • Shin, Hyun-Taeg;Lee, Kyung-Hee;Lee, Ah-Reum;Yoon, Seon-Hee
    • Journal of Korean Medicine for Obesity Research
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    • v.7 no.2
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    • pp.61-75
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    • 2007
  • Objectives : The purpose of this study was to investigate correlation between obese degree, result of hair tissue mineral analysis and bony maturation. And we also wanted to compare the differences between early bony maturation group(EBM) and late bony maturation group(LBM). Methods : 146 subjects who visited growth clinic were measured by BMI, PBF(percent body fat), percent BMI, bone age esimation and HTMA(hair tissue mineral analysis). The patients were classified into two groups - EBM, LBM group - according to the gap of bone age and chronological age. It was analysed that the correlation of bony maturation and obese degree, nutritional elements, heavy metals, significant ratio of nutritional elements. Also, analysed the differences between groups. Results : 1. BMI, percent BMI had a correlation with bony maturation, PBF however didn't have a correlation with it. 2. Ca, Mg, Zn and P had a positive correlation with bony maturation, and also K, Cr and Mo had a negative correlation. 3. U, As and Cd had a negative correlation with bony maturation. 4. Ca/P, Na/K, Ca/K and Zn/Cu had a positive correlation with bony maturation, and also Na/Mg, Ca/Mg had a negative correlation. 5. Percent BMI, Ca, Mg, Mn and Ca/p were higher in EBM group. 6. K, Mo and Hg were lower in EBM group. Conclusion : According to this study it could be suggested that maintaining proper percent BMI and accumulation of nutritional elements, heavy metals to prevent early bony maturation.

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High efficiency of homemade culture medium supplemented with GDF9-β in human oocytes for rescue in vitro maturation

  • Mohsenzadeh, Mehdi;Khalili, Mohammad Ali;Anbari, Fatemeh;Vatanparast, Mahboubeh
    • Clinical and Experimental Reproductive Medicine
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    • v.49 no.2
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    • pp.149-158
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    • 2022
  • Objective: Optimizing culture media for the incubation of immature oocytes is a vital strategy to increase the oocyte maturation rate during in vitro maturation (IVM) programs. This study evaluated the IVM and fertilization rates of human germinal vesicle (GV) and metaphase I (MI) oocytes using two different maturation media (commercial and homemade) with or without growth differentiation factor 9-β (GDF9-β). supplementation. Methods: Immature oocytes from intracytoplasmic sperm injection (ICSI) cycles were collected and assigned to one of two IVM culture media (commercial or homemade; cleavage-stage base). After maturation, MII oocytes were examined under an inverted microscope for the presence of the polar body, zona pellucida (ZP) birefringence, and meiotic spindle (MS) visualization after maturation in four conditions (commercial or homemade medium, with or without GDF9-β. ICSI was done for matured oocytes, and fertilization was confirmed by the visualization of two distinct pronuclei and two polar bodies. Results: No significant differences were found between the two culture media in terms of the time and rate of oocyte maturation or the rate of fertilization (p>0.05). Growth factor supplementation increased the 24-hour maturation rate for both GV and MI oocytes only in homemade medium. The maturation rate after 24 hours was higher for MI oocytes (p<0.05). Similar results were observed for MS visualization and ZP structure in both types of media (p>0.05). Conclusion: Higher rates of oocyte maturation and fertilization were observed after application of homemade medium supplemented with GDF9-β. Therefore, this combination may be recommended as an alternative for clinical IVM programs.

Effects of Sera, HEPES and Granulosa Cells Added to Culture Medium on In Vitro Maturation of Extrafollicular Bovine Oocytes (배지에 첨가한 혈청, HEPES 및 과립막세포가 난포외 소 난자의 체외성숙에 미치는 영향)

  • Hur Jun-Hoi;Hwang Woo-Suk;Jo Coons-Ho
    • Journal of Veterinary Clinics
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    • v.7 no.1
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    • pp.419-427
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    • 1990
  • Immature bovine oocytes were cultured to investigate whether the addition of FCS(10% or 20% ), CS (10%or 20% ) or BSA(5mg/ml) to culture medium with or without HEPES and co-culture with granulosa cells affect the frequency of in vitro maturation of extrafollicular bovine oocytes. After culture, the maturation rates were examined by the presence of 1st polar body and nuclear configuration. The maturation rate when FCS and CS as protein supplement were added to culture medium with or without HEPES was significantly higher than when BSA was added, and the maturation rate of extrafollicular bovine oocytes co-cultured with granulosa cells was higher than that cultured without granulosa cells, but there was no significant difference. FCS and CS were shown to be superior protein supplement when compared to BSA, and serum concentration, HEPES and co-culture with granulosa cells did not affect the in vitro-maturation of extrafollicular bovine oocytes.

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Changes in the Cell Wall Components, Enzyme Activities and Cell Structure of Peach during Maturation (복숭아의 품종과 숙도에 따른 세포벽 성분, 효소활성 및 조직의 변화)

  • 김미현;신승렬
    • Food Science and Preservation
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    • v.1 no.2
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    • pp.107-116
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    • 1994
  • This paper was carried out to Investigate changes in the activities of cell-degrading enzymes, cell wall components and cell structure of peach during maturation and storage for valuation of quality. The firmness of peach was decreased during maturation and storage, and was remarkably decreased in Daegubo than Yumyung. Polygalacturonase and $\beta$-galactosidase activities of peach were increased during maturation and storage, and were remarably increased in soft peach and in mature and soft peach, respectively. Contents of alcohol-insoluble substance, cell wall, and total and insoluble pectin of peach were decreased during maturation and storage, but cellulose and soluble pectin were increased. Intracellular space was enlarged during maturation and middle lamella was gradually degraded during maturation.

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Effect of Type and Culture Time of Porcine Oocytes On in Vitro Maturation and Developmental Potential of Embryos (돼지 난포란의 형태와 배양시간이 체외성숙 및 수정란의 배발생능에 미치는 영향)

  • 이장희;김창근;정영채
    • Journal of Embryo Transfer
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    • v.9 no.1
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    • pp.73-83
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    • 1994
  • The objective of this study was to develop an effective in vitro production system capable of obtaining more porcine embryos from immature oocytes. These experiments were thus conducted to examine the effect of oocytes type and maturation time on the in vitro maturation(IVM) and fertilization(IVF) of oocytes and the in vitro development (IVD)of IVF embryos. 1. The degree of oocyte maturation based on cumulus expansion index(GEI) did not differ for A- and B-typed oocytes but the index of oocyte type C was lower(P<0.05) than that of other oocyte types. 2. When the oocytes of type A and B were matured for 36, 42 and 48hrs, the GEl was not different between the 36- and 42-h maturation but the GEl after 48hrs was greatly lower(P<0.05) than that of other maturation times. 3. The highest cleavage rate(48.6%) of IVF oocytes was obtained from A typed oocytes and 42-h maturation but the developmental potential based on cleavage index was the highest when B-typed oocytes were matured for 42hrs.

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USEFULNESS OF CERVICAL VERTEBRAE MATURATION STAGE AS A MANDIBULAR MATURATION INDICATOR (하악골 성장 지표로서 경추골 성숙도의 유용성)

  • Choi, Bong-Sun;Choi, Nam-Ki;Kim, Sun-Mi;Yang, Kyu-Ho;Chung, Sung-Su
    • Journal of the korean academy of Pediatric Dentistry
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    • v.34 no.4
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    • pp.551-559
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    • 2007
  • This study was to investigate if cervical vertebrae maturation stages are as useful as hand-wrist maturation stages in evaluating the mandibular growth. The subject consisted of 292 girls aged from 8 to 16 years with normal occlusion. They were classified according to diagnosis by using studycast, lateral cephalogram, and handwrist X-ray film. The results were as follow: 1. Cervical vertebrae and hand-wrist maturation stages increased with age. 2. All mandibular measurements (Ar-Go, Go-Me, N-Go, S-Gn, N-Me) increased linearly with cervical vertebrae maturation stages. 3. Ar-Go, Go-Me, N-Go, S-Gn increased linearly with hand-wrist maturation stages. 4. Ar-Go, Go-Me, N-Go, S-Gn increased relatively rapidly between cervical vertebrae maturation stages 3 and 4. Go-Me and S-Gn increased relatively rapidly between hand-wrist maturation stages 6 and 7. 5. Ar-Go, Go-Me, N-Go, S-Gn, N-Me had high correlations with cervical vertebrae maturation stages as well as hand-wrist maturation stages. These results suggest that cervical vertebrae maturation stages are reliable on evaluating the mandibular growth.

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The Interaction between Epidermal Growth Factor (EGF) and Follicular Stimulating Hormone (FSH) on Nuclear Maturation of Mouse Oocytes by Using Their Inhibitor

  • Cha, Soo-Kyung;Kim, Tae-Hyung;Eum, Jin-Hee;Park, Kang-Hee;Park, Eun-A;Kim, Seung-Bum;Chung, Mi-Kyung;Lee, Dong-Ryul;Ko, Jung-Jae
    • Proceedings of the Korean Society of Embryo Transfer Conference
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    • 2002.11a
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    • pp.113-113
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    • 2002
  • The stimulatory effect of EGF and FSH on oocyte maturation have been reported in various mammalian species. And some reports presented FSH enhanced the effect of EGF on oocyte maturation. But, the interaction between EGF and FSH on nuclear maturation of mammalian oocytes is not fully understood. We observed the effect of EGF and FSH on nuclear maturation during in vitro maturation of mouse oocytes. Also, we examined the interaction between EGF and FSH on nuclear maturation of mouse oocytes using the EGFR inhibitor or FSH inhibitor. Germinal vesicle (GV) stage oocytes were obtained from 3-4weeks PMSG primed BCFI hybrid mice and cultured in TCM-199 medium with 0.4%PVP supplemented with/without EGF (1ng/ml), FSH (1ug/ml), EGFR specific tyrosine kinase inhibitors: Tyrphostin AG 1478 (500nM), MAP kinase kinase inhibitor : U0126 (20uM) or PD 98059 (100uM) for 14-l5hr. Rapid staining method were used for the assessment of nuclear maturation. Nuclear maturation rates of EGF indjor FSH-treated group were significantly higher than those of control group. Treatment of EGFR inhibitor significantly block the nuclear maturation of GV oocyte in EGF-treated group, but it did not block those of GV oocyte in FSH-treated or FSH and EGF-treated group. Treatment of FSH inhibitor(U0126, PD98059) significantly block the nuclear maturation of EGF-treated group, FSH-treated and FSH and EGF-treated group. These results show that EGF has a stimulatory effect as well as different action pathway with FSH on in-vitro maturation of mouse oocyte in vitro. Therefore, further studies will be needed to find the signaling pathway of EGF associated with nuclear maturation.

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The Effects of Transcription / Translation Inhibitors on Meiotic Maturation of Porcine Oocyte In Vitro

  • Byun, Tae-Ho;Lee, Sung-Ho;Park, Chang-Sik;Lee, Sang-Ho
    • Proceedings of the Korean Society of Embryo Transfer Conference
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    • 2002.11a
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    • pp.117-117
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    • 2002
  • The oocytes from most of animal species accumulate genetic information and other necessary materials during oogenesis for the later use in the early development. Over the years oocyte maturation has been studied extensively both in vitro and in vivo. Particularly, maturation of follicular oocyte in vitro becomes one of the important tools for the studies of basic cell biology, the in vitro technology of animal production, and in particular, the somatic cell cloning by nuclear transfer. We examined meiotic maturation and cumulus expansion in the presence of translation or transcription inhibitors for varying periods of in viかo maturation (IVM) of pig oocyte. In Experiment 1, the results revealed that translation and transcription inhibitors inhibited cumulus expansion and meiotic maturation during 35h of IVM. However, 50 to 60% of the oocytes underwent nuclear maturation without cumulus expansion during 75h of IVM. The rest of the oocytes were arrested at metaphase I (40-50%) in the presence of the inhibitors. In Experiment II, the OCCs were exposed to the drugs only for 15h to examine translation and transcription inhibitors on cumulus expansion and meiotic maturation. Transcription inhibitors for 15h did not arrest meiotic maturation when the oocytes were cultured for subsequent, necessary period of IVM, whereas cumulus expansion was completely inhibited, suggesting that initial 15h is critical transcription activity far cumulus expansion. Translation inhibitors for 15h exposure did not alter cumulus expansion and meiotic maturation during subsequent culture in the absence of the drugs. In Experiment III, the OCCs were exposed to the drugs only for later 30h to examine the influence of transcription and translation inhibitors on oocyte maturation. Interestingly, all meiotic maturation underwent normally with full expansion of cumulus. Similar results were obtained from Experiment IV where 5h of exposure from 15 to 20h of IVM culture to the drugs was performed and subsequently cultured for same period in fresh medium. Taken there results together, both transcription and translation are necessary for nuclear maturation and cumulus expansion, and first 15h IVM for cumulus expansion is critical. The arrested oocytes by the drugs were still capable of undergoing nuclear maturation, although cumulus expansion was affected.

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Redistribution of Intracellular $Ca^{2+}$ Stores during Mouse OOcyte Maturation (생쥐 난자 성숙시 일어나는 칼슘 저장고의 분포 변화에 관한 연구)

  • 최수완
    • Development and Reproduction
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    • v.1 no.1
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    • pp.45-56
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    • 1997
  • Befor fertilization, mammalian oocytes undergo meiotic maturation, which consists of nuclear and cytoplasmic differentiation. In this study, changes of $Ca^{2+}$ stores in mouse oocytes were examined during meiotic maturation and the role of $Ca^{2+}$ in the regulation of the maturation was investigated by using monoclonal antibodies against smooth endoplasmic reticulum $Ca^{2+}$-ATPase(SERCA-ATPase) and calreticulin. Observations were made under epifluorescence microscope and/or confocal laser scanning microscope. In immature oocytes which did not resume meiotic maturation, SERCA-ATPases were mostly localized in the vicinity of the germinal vesicle and calreticulins were distributed evenly throughout the cytoplasm. In mature oocytes, SERCA-ATPases were observed throughout the cytoplasm, butwere absent from the nuclear region. In contrast, calreticulins were localized mostl in the cortex of the oocyte and were absent from the cytoplasm. However, bright fluoresence stainings were wbserved in the perimeiotic spindle region of mature oocyte when labeled with antibodies against calreticulin. These results indicate that mouse oocytes undergo distinct rearrangement of the localization of $Ca^{2+}$-ATPases and calreticulins during meiotic maturation. Thus it can be suggested that redistribution of the $Ca^{2+}$ stores, as revealed by differential fluorescence stainings, is deeply involved in the regulatory mechanism of mammalian oocyte maturation.

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