• Title/Summary/Keyword: maturation

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Studies on In Vitro Fertilization and Development of Bovine Follicular Oocytes Matured In Vitro V. Effect of Hyaluronic acid,h Chondroitin Sulfate, and Heparin on In Vitro Maturation of Bovine Follicular Oocytes (체외성숙 우난포란의 체외수정과 발달에 관한 연구 V. Hyaluronic acid와 Chondroitin Sulfate 및 Heparin이 우난포란의 체외성숙에 미치는 영향)

  • 박세필;정형민;오종훈;이훈택;정길생
    • Korean Journal of Animal Reproduction
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    • v.16 no.1
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    • pp.7-14
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    • 1992
  • These experiments were designed to investigate the effects of glycosaminoglycans(GAGs), which maybe produced by cumulus cells, on in vitro maturation process of bovine follicular oocytes. The rates of in vitro maturation of bovine oocytes were examined after incubating with the various concentrations of a hyaluronic acid, chondroitin sulfate, or heparin for 26 hours. The results obatained in these experiments were summarized as follows : 1. When the cumulus or removed oocytes treated with hyaluronic acid(200, 400, 800 and 1, 600$\mu\textrm{g}$/ml), the maturation rates of cumulus-removed oocytes were 48.6, 59.4, 68.4 and 61.3% respectively. Especially, at a concentration of 800$\mu\textrm{g}$/ml, the maturation rates(68.8%) of cumulus-removed oocytes were slightly lower than that(78.3%) of cumulus-enclosed oocytes. However, the treatment of hyaluronic acid showed significantly higher maturation rates compared to that(48.4%) of control group of cumulus-removed oocytes(p<0.05). Therefore, these results suggest thate hyaluronic acid had a beneficial effect on maturation of bovine follicular oocytes. 2. The maturation rate of cumulus-free oocytes treated with chondroitin sulfate(200, 400, 800 and 1, 600$\mu\textrm{g}$/ml) was 52.3, 56.1, 55.9 and 52.2% respectively. These rates were not different from those(52.9%) of control group. However, these rates in cumulus-free oocytes were significantly lower than that(79.0%) in cumulus-enclosed oocytes(p<0.01). Therefore, these results suggest that chondroitin sulfate do not affect on the maturation of oocytes. 3. In cumulus-free oocytes cultured with different heparin concentration, the maturation rates were ranged from 48.5 to 52.1%, showing no differencies from that(50.7%) of control group. However, these rates were significantly lower than 80.0% in cumulus-enlosed oocytes(p<0.01). 4. The nuclear maturaton of oocytes was increased by treatment of each of three glycosaminoglycans(hyaluronic acid, chondroitin sulfate, and heparin). In addition, the treatment of mixed together showed the significant additive effect. Hyaluronic acid was more effective than chondroitin sulfate and heparin were.

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Effect of Epidermal Growth Factor on In Vitro Maturation in Pig Immature Oocytes I. Effect of Epidermal Growth Factor in Nuclear Maturation (Epidermal Growth Factor가 돼지 미성숙난포란의 체외성숙에 미치는 영향 I. 핵성숙에 미치는 Epidermal Growth Factor의 효과)

  • 엄상준;김선의;김은영;윤산현;박세필;정길생;임진호
    • Korean Journal of Animal Reproduction
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    • v.19 no.3
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    • pp.217-226
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    • 1995
  • The objective of this experiment was to test the effect of EG F on nuclear maturation of pig immature oocytes in vitro. Basic medium used TCM-199 supplemented with 0.2 mM pyruvate, 1 ${\mu}\textrm{g}$/ml estradiol-I7$\beta$ and 25 ${\mu}\textrm{g}$/ml gentamycin, this medium treated with EGF, FSH and FBS. Experiment 1 examined to the effect according to the addition of FSH or EGF (0, 1. 10 and 100 ng EGF/ml) in oocytes maturation. Nuclear maturation rates (M ll %) of 1, 10 and 100 ng EGF/ml (83.0. 8fi.7 and H7.5%) treatments were significantly higher than those of non- and FSH-treated groups (27.3 and 60.3%, p < 0. 001). Experiment 2 examined to the interactive effects of EGF. FSH or FBS during oocytes maturation. Nuclear maturation rates (M ll %) of EGF alone, EGF plus FSH, EGF plus FBS, FSH plus FBS, and EGF plus FSH added FBS treatments (86.7, 90.2, 87.1. 89.6% and 92.6%) were significantly higher than those of non, FSH, and FBS alone treatments (22.3, 52.2 and 42.3%, p < 0.001). Also, cumulus cells expansion of oocytes maturation was examined to total treatments. Normal cumulus cells expansion was shown by FSH plus FBS, EGF or EGF with FBS combination treatments, but cumulus cells of oocyte complexes were still clumped together in EGF-treated groups although they had separated from oocytes. However, EGF showed a positive on nuclear maturation. These results conclude that EGF alone can stimulate nuclear maturation in pig immature oocytes.

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Effect of Epidermal Growth Factor and Transforming Growth Factor-$\alpha$ on In Vitro Maturation of Porcine Oocytes (Epidermal Growth Factor(EGF)와 Transforming Growth Factor-$\alpha$(TGF-$\alpha$)가 돼지 난포란의 체외성숙에 미치는 영향)

  • 임정훈;박병권;이규승
    • Korean Journal of Animal Reproduction
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    • v.21 no.2
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    • pp.177-183
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    • 1997
  • The present study examined the effects of epidermal growth factor(EGF) and transforming growth factor-$\alpha$(TGF-$\alpha$) on in vitro maturation of porcine follicular oocytes. Basic medium used TCM-HEPES, and oocytes cultured for 42 hours in vitro. The results obtained are as follows; 1. The nuclear maturation rates of EGF-treated groups(10ng/ml, 75.9% ; 30ng/ml, 69.2% ; 50ng/ml, 67.2% ; 100ng/ml, 71.0%) on the porcine oocytes cultured in medium without pFF in vitro were significantly(P<0.01) higher than those of non-treated group(57.1%). When the oocytes were cultured in media with 10%(v/v) pFF, the nuclear maturation rates of 30ng EGF/ml(77.1%) treated group were significantly(P<0.01) higher than those of non-(59.2%) and EGF-treated groups(10ng/ml, 65.4% ; 50ng/ml, 65.5% ; 100ng/ml, 70.4%). 2. The nuclear maturation rates of 30ng TGF-$\alpha$/ml treated group(71.9%) in media without pFF in vitro were significatnly(P<0.01) higher than those of non-(57.1%) and TGF-$\alpha$ treated groups(10ng/ml, 60.4% ; 50ng/ml, 65.4% ; 100ng/ml, 60.0%). When the oocytes were cultured in media with 10%(v/v) pFF, the nuclear maturation rates of 30 and 50ng TGF-$\alpha$/ml(77.4% and 79.6%) treated groups(10ng/ml, 64.2% ; 100ng/ml, 61.6%). 3. On the effect of EGF(30ng/ml) and/or TGF-$\alpha$(30ng/ml) treated groups in medium without pFF in vitro, the nuclear maturation rates indicated 57.3, 60.4, 75.9 and 79.7% in media with no EGF & TFG-$\alpha$, TGF-$\alpha$ only, EGF only nad EGF+TGF-$\alpha$ treated groups, respectively. The nuclear maturation rates in medium with EGF only or EGF+TGF-$\alpha$ were significantly(P<0.01) higher than those non- and TGF-$\alpha$ treated groups. When the oocytes were cultured in media with 10%(v/v) pFF, the nuclear maturation ratesof EGF+TGF-$\alpha$ treated group(75.9%) were significantly(P<0.01) higher than those of non-(59.2%), TGF-$\alpha$ only (64.2%) and EGF only(69.4%) treated groups.

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Influences of Human Body Fluids and Gonadotropins Supplemented in the Maturation Medium on the Nuclear Maturation and Fertilizability of Mouse Immature Oocytes (성숙배양액에 첨가하는 인간체액 (Human Body Fluids) 및 성선자극호르몬이 생쥐 미성숙난자의 핵성숙과 수정능력에 미치는 영향)

  • Park, K.S.;Son, W.Y.;Kim, J.H.;Lee, K.A.;Han, S.Y.;Ko, J.J.;Cha, K.Y.
    • Clinical and Experimental Reproductive Medicine
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    • v.21 no.2
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    • pp.183-190
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    • 1994
  • Purpose of the present study was to find the optimal culture conditions for the maturation and fertilization of immature oocytes by the use human body fluids and gonadotropins (Gn) in the mouse model. Cumulus-enclosed mouse immature oocytes were incubated in the medium containing various human body fluids with or without Gn in vitro, and examined to confirm nuclear maturation (NM) and fertilization. Female ICR mice were stimulated with 7.5 IU pregnant mares' serum gonadotropin (PMSG). Cumulus-enclosed immature oocytes were isolated at 48-52 hr post PMSG injection and cultured in TCM 199 supplemented with various concentrations (20, 50, and 70%) of human body fluids such as fetal cord serum (hCS), follicular fluid (hFF), peritoneal fluid (hPF) and amniotic fluid (hAF) in the presence or absence of 10 IU/ml PMSG and 10 IU/ml human chorionic gonadotropin (hCG) for 18 hr. Fetal calf serum (FCS) was used as a control for the supplements. Matured oocytes were fertilized with sperm collected from the epididymis of male mice. Fertilization was conducted in T6 medium containing 15 mgl ml bovine serum albumin, and confirmed at 6 hr post-insemination. Evaluation of nucler maturation and fertilization was carried out by rapid staining using fuchin. There was no significant difference between the effects of human body fluids and FCS supplements on nuclear maturation of cumulus enclosed mouse immature oocytes. When maturation medium was supplemented with 20% hPF or 20% hAF, fertilization rates were significantly (P<0.01) lower than that of 20% FCS, hCS and hFF groups. However, higher concentrations of body fluids during IVM were not more beneficial on fertilizability of oocytes. The addition of Gn significantly increased the fertilization rates in hPF and hAF groups (hPF without Gn; 51.5%, compared with 85.1% for addition of Gn, and hAF without Gn; 30.1% compared with 85.8% for addition of Gn) at 20% concentration. These results suggest that human body fluids at 20% concentration and gonadotropins can be used as supplements for the maturation of mouse immature oocytes in vitro. When gonadotropins supplemented with the human body fluids in the maturation medium, fertilizability of mouse immature oocytes was increased in hPF and hAF groups. These results can be applied to maturation of human immature oocytes in vitro.

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Interaction between Transforming Growth Factor $\beta$ and Cumulus Cells during In Vitro Maturation in Porcine Oocytes (돼지난자의 체외성숙시 Transforming Growth Factor$\beta$와 난구세포의 상호작용)

  • 신명균;조재원;정희태;양부근;김정익;박춘근
    • Korean Journal of Animal Reproduction
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    • v.22 no.1
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    • pp.73-80
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    • 1998
  • This study was undertaken to evaluate the interaction between cumulus cells and TGF $\beta$1 on in vitro maturation in porcine oocytes. No differ ences were found in maturation rates when follicular oocytes were cultured in medium with various concentrations of TGF $\beta$. At 24 h after maturation, the oocytes matured to metaphase-II were found in medium with TGF $\beta$ regardless of cumulus cells. On the other hand, the maturation rates were significantly(P < 0.01 higher cumulus-enclosed(70 and 52%) than cumulus-denuded oocytes(35 and 26%) in medium with or without TGF $\beta$ at 48 h after culture. In a another experiment, the same maturation rates (54-71%) were observed when cumulus-enclosed oocytes were cultured with various addition time of TGF $\beta$. However, the maturation rates in cumulus-denuded oocytes were significantly (P < 0.05) higher in medium added at 0~24 h (59%) or 24-48 h(57%) after culture than in medium with(27%) and without(38%) TGF $\beta$ for 48 h. These results indicated that cumulus cells is essential for in vitro maturation in porcine oocytes but TGF $\beta$ can promote oocytes maturation in cumulus-free oocytes.

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Nuclear Maturation and Pronuclei Formation in Bovine Oocytes Matured In Vitro for Prolonged Period (체외 성숙 시간에 따른 소 난자의 처녀 발생)

  • 유형진;최승철;이상호
    • Korean Journal of Animal Reproduction
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    • v.17 no.4
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    • pp.331-337
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    • 1994
  • Response of the oocytes to parthenogenetic activation is one of the indice for cytoplasmic maturation. Maturational age-dependent parthenogenetic activation was examined in bovine oocytes. Follicular oocytes recovered from the slaughter house ovaries were matured in vitro in TCM 199+15% FCS+1Oiu/ml PMSG +10 iu/ml hCG from 24 to 48 h at 6 h intervals. The in vitro matured oocytes were activated by 7% ethanol for 7 min. The nuclear maturation and the cytoplasmic maturation were analysed by the nuclear configuration and pronuclei formation stained by rapid staining method. Cumulus oophori expansion increased as the maturation time increased. Proportions of the nuclear maturation were 81, 89, 72, 60 and 60% in IVM 24, 30, 36, 42 and 48 h groups, respectively. Abnor¬mality in metaphase II chromosome increased sharply from 36 h IVM. The rates of the pronuclei formation and diploid upon ethanol activation were 67, 68, 73, 84 and 87%, and 4, 5, 10, 16 and 20% in IVM 24, 30, 36, 42 and 48 h groups, respectively. It was suggested that maturational age increased the formation of the pronuclei and diploid, and that cytoplasmic maturation require longer maturation period than normal nuclear maturation. These results should be useful for determination of an appropriate time for fertilization in mammalian eggs matured or preincubated in vitro.

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Assessment of Midpalatal Suture Maturation by Skeletal Maturity on Hand Wrist Radiographs (성장기 아동의 수완부 골 성숙도를 이용한 정중구개봉합 성숙도 평가)

  • Yu, Dayeol;Kim, Donghyun
    • Journal of the korean academy of Pediatric Dentistry
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    • v.48 no.1
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    • pp.31-41
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    • 2021
  • The aim of this retrospective study was to evaluate the correlation between the midpalatal suture maturation and skeletal maturation in growing children aged 7 - 15 years and predict the maturational stages of the midpalatal suture corresponding to skeletal maturity assessed by the skeletal maturation indicators (SMI) and middle phalanx of the third finger (MP3) stages. The group of this retrospective study was consisted of randomly selected 132 male and 135 female in age from 7 - 15 years. The maturation of the midpalatal suture was evaluated by using images from cone-beam computed tomography (CBCT) while the skeletal age was assessed by hand-wrist radiography. CBCT images and hand-wrist radiographs used in this study were obtained from all subjects for orthodontic diagnosis before orthodontic treatment. The maturational stages of the midpalatal suture showed strong correlations with both SMI and MP3 stages. The correlation between the midpalatal suture maturation and SMI (Spearman's correlation coefficient, ϒS = 0.905, p < 0.05) was slightly greater than that of MP3 stages (ϒS = 0.830, p < 0.05). There was a positive significant correlation between the midpalatal suture maturation and chronological age (ϒS = 0.868,p < 0.05). CBCT for evaluation of the midpalatal suture maturational stages may be unnecessary in every pediatric patients because SMI and MP3 stages were both replaceable useful methods for assessing maturation of the midpalatal suture before orthopedic treatment. In this retrospective study, the diagnostic reliability of the SMI method for estimating midpalatal suture maturation showed better reliability than the MP3 method.

Effect of Insulin-like Growth Factor-I and Cumulus Cells on In Vitro Maturation in Porcine Oocytes (돼지난자의 체외성숙시 Insulin-like Growth Factor-과 난구세포의 영향)

  • Park, C.K.;Cho, J.W.;Shin, M.K.;Cheong, H.T.;Yang, B.K.;Kim, C.I.
    • Korean Journal of Animal Reproduction
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    • v.23 no.4
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    • pp.323-331
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    • 1999
  • The effects of insulin-like growth factor-I (IGF-I) and cumulus cells during in vitro maturation in porcine oocytes were examined. When follicular oocytes were cultured in medium with different concentrations of IGF-I, maturation rates were 60, 61 and 62 and 72% for 0, 15 and l0ng/$m\ell$ IGF-I. In medium with 10ng/$m\ell$ IGF-I, maturation rates were not significantly difference between oocytes with (68%) and without (52%) cumulus cells during the culture. In medium with-out IGF-I, however, the maturation rates in oocytes with cumulus cells (63%) was significantly (P<0.05) higher than oocytes without cumulus cells (32%). On the other hand, when IGF-I was added for first 24 h period or later 24 h period of culture, maturation rates were higher in oocytes with (61 and 49%) that than without (49 and 45%) cumulus cells. In experiment used medium without fetal calf serum (FCS) and porcine follicular fluid (PFF), the maturation rates in oocytes with cumulus cells for 48 h (48 and 67%) or first 24 h (46 and 63%) period after culture were significantly (P<0.01) higher than in oocytes without cumulus cells (16 and 18%) in the presence or absence of IGF-I. These results indicated that cumulus cells is essential on maturation in vitro in porcine oocytes, but IGF-I can promote oocytes maturation of oocytes without cumulus cells in medium with FCS and PFF.

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Heat waves impair cytoplasmic maturation of oocytes and preimplantation development in Korean native cattle (Hanwoo)

  • Sa, Soo Jin;Jeong, Jiyeon;Cho, Jaesung;Lee, Seung-Hwan;Choi, Inchul
    • Korean Journal of Agricultural Science
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    • v.45 no.3
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    • pp.493-498
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    • 2018
  • There has been widespread warming and a general increase in summer temperatures over the Korean peninsula ($0.5^{\circ}C$/10 years from 2001 to 2010). South Korea is transforming into a subtropical region, and the productivity of livestock is affected by the climatic changes. In this study, we investigated whether the summer heat waves affect the developmental competency of Korean native cattle (Hanwoo), a taurine type of cattle with a small portion of indicine varieties. We collected oocytes during the summer (heat stress, HS) and autumn (non-HS condition) and examined the developmental competencies including in vitro maturation and preimplantation embryo development. No significant differences were observed between the HS and non-HS oocytes in nuclear maturation (extrusion of the polar body); however, the cleavage and blastocyst rates were significantly lower in the HS group than those in the non-HS group. The lower developmental competence of the HS oocytes compared to the non-HS is, in part, due to insufficient cytoplasmic maturation because of a higher production of Reactive oxygen species (ROS) levels as well as peri/cortical distributed mitochondria in the HS oocytes after in vitro maturation. Next, we examined the ROS and mitochondria distribution and found a significant increase in the levels of ROS in the HS oocytes and a polarized distribution (pericortical cytoplasm) of mitochondria in the HS oocytes. In summary, impaired cytoplasmic maturation of oocytes from exposure to HS affects the preimplantation embryo development by dysfunction of mitochondria. To improve reproductive performance, embryo transfer using cryopreserved embryos/oocytes is recommended in the hot summer season of South Korea.

The Effect of Reduction of Contaminants and Odor according to the Additives in the Anaerobic Maturation Process of Piggery Slurry (양돈장 분뇨의 혐기성 부숙 과정에서 첨가제에 따른 오염물질 및 악취 저감 효과)

  • Kang Kyung-Ho;Kam Sang-Kyu;Hu Chul-Goo;Lee Min-Gyu
    • Journal of Environmental Science International
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    • v.15 no.2
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    • pp.169-175
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    • 2006
  • The changes of contaminants and odor corresponding to anaerobic maturation process of piggery slurry were investigated by applying the additives, such as different kinds of complex microorganism products and deodorants containing microorganism activating agents. The pHs during 20-day anaerobic maturation were operated stably without great change regardless of the additives, although they were rather lower in the case that the additives were contained than the case that they were not contained. The effects of removing CODcr, $NH_3-N$, T-N, and T-S in case that the additives were not contained, were not so great during the 20-day operation and so it would be difficult to remove the organic materials and nitrogen ingredients simply with anaerobic maturation process. However, in case of anaerobic maturation process that the additives were contained, their average removal rates were improved with the values of $49\%,\;63.5\%,\;48.5\%,\;and\;30.7\%$ for above each of items, even if the 20-day of short-term maturation period was applied. Especial1y, odor intensity with the additives was lowered continuously during the operation period and it had more than two times of lowering effect compared to that without those.