• 제목/요약/키워드: Developmental rates

검색결과 617건 처리시간 0.027초

한우 체외수정란을 이용한 핵 이식배의 체외발달에 관한 연구 (In Vitro Development of Nuclear Transplantation Bovine Embryos Using In Vitro Fertilized Embryos of Korean Native Heifers)

  • 박충생;공일근;노규진;이효종;최상용
    • 한국가축번식학회지
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    • 제18권2호
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    • pp.113-119
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    • 1994
  • To improve nuclear transplantation(NT) efficiency and to produce a large scale genetically identical cloned calves, examined the in vitro development capacity after co-culture of bovine oviductal epithelial cells (BOEC) and granulosa cells in TCM-199 supplemented with 10% fetal calf serum (FCS) with early bovine embryos derived from in vitro matured fertilized(IVM-IVF) oocyte. In addition, the age dependence of IVM oocyte on electro-stimulation and the effective electric voltage on in ivtro development of bovine NT embryos were examined. The results obtained were summerized as follows; 1. The cleavage rates of IVM-IVF bovine embryos in co-culture with bovine oviductal epithelial cells and granulosa cells were not significantly different(P<0.05), but the developmental rate into morula and blastocyst stage were different showing 38.3 and 20.2%, respectively. 2. The activation (82.5%) and development in vitro(8.6%) into later embryo stages of the aging oocytes of 32 hours post-maturation (hpm) were significantly higher than those of 24 hpm at direct current (DC) voltage of 1.5kV/cm, 60$\mu$sec pulse duration and 1 pulse time. 3. The fusion rates of NT eggs of 32 hpm following to different DC voltages from range 0.75 to 1.5kV/cm were not differ, but the developmental rate into morula and blastocyst stages at DC voltages of 0.75 and 1.0kV/cm were higher(11.4 and 12.6%, respectively) than those of 1.5kV/cm(0%). From these results, it can be suggested the optimal culture system for in vitro culture of IVM-IVF bovine embryos is a co-culture system with BOEC in TCM-199 supplemented 10% FCS. The effective time and the DC voltage for activation, electrofusion and in vitro development of NT embryos derived from IVM-IVF bovine embryo are 32hpm and 0.75~1.0kV/cm. But to improve NT efficiency, the advanced research (cell cycle synchronization, micromanipulation, culture system, etc.) is needed.

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Changes in the Expressional Levels of Sarcoplasmic Reticulum $Ca^{2+}-regulatory$ Proteins in the Postnatal Developing Rat Heart

  • Lee, Eun-Hee;Park, Soo-Sung;Lee, Jae-Sung;Seo, Young-Ju;Kim, Young-Hoon;Kim, Hae-Won
    • The Korean Journal of Physiology and Pharmacology
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    • 제6권2호
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    • pp.101-107
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    • 2002
  • In the present study, the postnatal developmental changes in the expressional levels of cardiac sarcoplasmic reticulum (SR) $Ca^{2+}$ regulatory proteins, i.e. $Ca^{2+}-ATPase,$ phospholamban, and $Ca^{2+}$ release channel, were investigated. Both SR $Ca^{2+}-ATPase$ and phospholamban mRNA levels were about 35% of adult levels at birth and gradually increased to adult levels. Protein levels of both SR $Ca^{2+}-ATPase$ and phospholamban, which were measured by quantitative immunoblotting, were closely correlated with the mRNA levels. The initial rates of $Ca^{2+}$ uptake at birth were about 40% of adult rates and also increased gradually during the myocardial development. Consequently, the relative phospholamban/$Ca^{2+}-ATPase$ ratio was 1 in developmental hearts. $Ca^{2+}$ release channel (ryanodine receptor) mRNA was about $50{\sim}60%$ at birth and increased gradually to adult level throughout the postnatal rat heart development. $^3[H]ryanodine$ binding increased gradually during postnatal myocardial development, which was closely correlated with ryanodine mRNA expression levels during the development except the ryanodine mRNA level at birth. These findings indicate that cardiac SR $Ca^{2+}-ATPase,$ phospholamban, and $Ca^{2+}$ release channel are expressed coordinately, which may be necessary for intracellular $Ca^{2+}$ regulation during the rat heart development.

Fluorescence in situ hybridization(FISH) 기법을 이용한 인간 생식세포 및 착상전 배아의 유전이상 검색 (Detection of genetic abnormalities in human sperm, oocytes, and preimplantation embryos using fluorescence in situ hybridization (FISH))

  • 방명걸
    • 한국발생생물학회:학술대회논문집
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    • 한국발생생물학회 1998년도 제4차 학술발표대회 및 정기총회
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    • pp.12-18
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    • 1998
  • Tremendous progress has been made over the past quarter-century studying the genetics of gametogenesis and the resulting gametes and embryos. Studies merging molecular techniques and conventional cytogenetics are now beginning to bridge the gap between what we have learned about the meiotic process in males and females and what we know of the mitotic chromosomes of zygotes. Numerical abnormalities in sperm, oocytes and embryo can now diagnosed by fluorescence in situ hybridization (FISH). "At risk" couples can, therefore, have only unaffected embryos replaced in the sterus and avoid the possibility of terminating a pregnancy that might only be diagnosed as affected later gestation. Single-cell genetic analysis has also provided powerful tools for studying genetic defects arising during early human development. Recent studies of sperms, oocytes and cleavage-stage human embryos have revealed an unexpectedly high incidence. These genetic abnormalities are likely to contribute to early pregnancy loss and have important implications for improving pregnancy rates in infertile couples by assisted reproduction. The widespread use of preimplantation genetic diagnosis (PGD) awaits further documentatio of safety and accuracy. Other issues also must be addressed. First, the ethical issues regarding germ cell and embryo screening must be addressed including what diseases are serious enough to warrant the procedure. Another concern is the use of this technology for non-genetic disorders such as gender selection. Finally, the experimental nature of these procedure must continually be discussed with patients, and long-term follow-up studies must be undertaken. Development of more accurate and less expensive assays coupled with improved assisted reproductive technology success rates may make PGD a more widely use clinical tool. The future awaits these development.velopment.

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돼지 난자의 체외성숙과 배아발달 동안 ROS와 항산화제의 영향 (Effects of Reactive Oxygen Species and Antioxidants during In Vitro Maturation Oocytes and Embryo Development in Pigs)

  • 이원희;박지은;황보용;김화영;이지은;강병범;정희태;양부근;박춘근
    • Reproductive and Developmental Biology
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    • 제41권1호
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    • pp.17-23
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    • 2017
  • The oocyte undergoes various events during in vitro maturation (IVM) and subsequence development. One of the events is production of reactive oxygen species (ROS) that is a normal process of cell metabolism. But imbalances between ROS production and antioxidant systems induce oxidative stress that negatively affect to mammalian reproductive process. In vitro environments, in vitro matured oocytes have many problems, such as excessive production of ROS and imperfect cytoplasmic maturation. Therefore, in vitro matured oocytes still have lower maturation rates and developmental competence than in vivo matured oocytes. In order to improve the IVM and in vitro culture (IVC) system, antioxidants, vitamins were added to the IVM, IVC medium. Antioxidant supplementation was effective in controlling the production of ROS and it continues to be explored as a potential strategy to overcome mammalian reproductive disorders. Based on these studies, we expect that the use of antioxidants in porcine oocytes could improved maturation and development rates.

Nuclear Remodeling and In Vitro Development Following Somatic Cell Nuclear Transfer in Swine

  • Yoon Jong-Taek;Kim Yong-Yeup;Lee Jong-Wan;Min Kwan-Sil;Hwang Seongsoo
    • Reproductive and Developmental Biology
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    • 제28권4호
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    • pp.241-245
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    • 2004
  • This study was conducted to investigate nuclear remodeling and developmental rate following nuclear transfer of fetal fibroblast cells, ear skin cells and oviduct epithelial cells into porcine recipient oocytes. To test par-thenogenetic activation, oocytes were treated with a 6-dimethylaminopurine (6-DMAP), a single DC-pulse (DC), calcium ionomycin (ionomycin), DC+6-DMAP and ionomycin + 6-DMAP after in vitro maturation. For nuclear transfer, in vitro matured oocytes were enucleated, and donor cells were transferred into oocytes. Cloned embryos were fused and stimulated with 6-DMAP for 4 h and cultured in vitro for 6 days. Among treatments for parthenogenesis, the activation rate of DC +6-DMAP treatment was significantly higher than that of single treatment roups (p<0.01), except for DC treatment group. However, the difference was not significant in activation rate compared to other complex treatment groups. Nuclear swelling of the cloned embryos was initiated at 60 min after stimulation and increased afterwards. Fusion rates were not different among different donor cells. Cleavage rates of DC treatment groups were significantly higher than those of DC+6-DMAP treatment groups (p<0.05) in case that fetal fibroblast and ear cells were used for nuclear donor. The cloned embryos from developed to blastocysts in oviduct epithelial cell nuclear transfer with DC+6-DMAP treatment was significantly higher compared to those with DC only treatment (p<0.05). However, no blastocyst was developed from nuclear transfer of fetal fibroblast and ear cells regardless of activation treatments. Based on these results, a proper activation stimulation may be necessary to increase the activation rate and the development to blastocyst in cloned porcine embryos.

Epidermal Growth Factor(EGF)가 생쥐 초기배아의 발생에 미치는 영향 (Effect of Epidermal Growth Factor(EGF) on Early Embryonic Development in Mouse)

  • 변혜경;이호준;김성례;김해권;김문규
    • Clinical and Experimental Reproductive Medicine
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    • 제22권2호
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    • pp.163-170
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    • 1995
  • Growth factors (GFs) produced by the embryo or by the maternal reproductive tract have been reported to regulate the embryonic development and differentiation. Among GFs, EGF as a mitogen plays a role in mitosis and functional differentiation of trophectoderm cells in mouse. The present study was carried out to investigate the effect of EGF on development of mouse embryos and to localize EGF in the mouse oocytes and embryos, which has been reported to be detected in the reproductive tract in mammals. To investigate the effect of EGF on the development of the embryo, mouse 2-cell embryos were cultured to blastocysts stage in Ham's F10 medium, treated with EGF(10-50 ng/ml) for 72 hrs. Immunocytochemistry was performed from oocyte to blastocyst stage with anti-EGF and anti-Mouse IgG, in order to determine the stage which EGF would be expressed in mouse. Exogenous EGF (more than 10 ng/ml) in the culture medium improved the developmental and hatching rates in the mouse embryos. As a result of immunocytochemistry, the embryonic EGF was expressed after the late 4-cell stage. EGF is thought to enhance preimplantation embryonic development and hatching. Exogenous EGF in the culture medium is thought to activate EGF receptor in the late 4-cell embryos and to enhance blastulation and hatching in mouse embryos. It is concluded that EGF enhances the developmental and hatching rates in the mouse embryos.

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Vitrification 방법에 의한 토끼수정란의 동결에 관한 연구 (Cryopreservation of rabbit embryos by vitrification)

  • 최상용;이영락;노규진;이효종;박충생
    • 대한수의학회지
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    • 제35권3호
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    • pp.635-641
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    • 1995
  • The purpose of this study was to investigate the effects of developmental stage and equilibration time on survival of rabbit embryos following freezing by vitrification. Adult New Zealand White female rabbits were superovulated with PMSG and hCG. The 8-cell stage embryos were collected from 40 to 45 hours after hCG injection by flushing oviducts with Dulbecco's phosphated buffered saline and in vitro cultured in TCM-199 containing 10% fetal calf serum(FCS). Each embryos developed in vitro to 16-cell, compact morula and blastocyst was cryopreserved and cultured following thawing to examine their developmental potential to expanded blastocyst stage in vitro. The frozen-thawed-cultured embryos were stained with Hoechst 33342, and their nuclei were counted using a fluorescence microscope. On the toxicity test of EFS solution as cryopreservation, the survival rates of 8-cell stage embryos was decreased in reverse to increasing of exposure time over 5 minutes. The post-thaw survival rates of embryos on equilibration times was significantly(P<0.05) higher for 2 min. than for 5 or 10 minutes. From morula to blastocyst of rabbit embryos was more suitable than 8-cell stage for cryopreservation by vitrification. The higher post-thaw survival rate of embryos can be achieved by keeping the cryoprotectant at $4^{\circ}C$ than at $20^{\circ}C$. The mean number of nuclei per embryo following freezing by vitrification and in vitro culture to expanded blastocyst at compacted morula and blastcyst was not significantly differ from fresh blastocyst.

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Blastocyst formation in vitrified-warmed preimplantation embryos derived from vitrified-warmed oocytes in a mouse model

  • Yeon Hee Hong;Byung Chul Jee
    • Clinical and Experimental Reproductive Medicine
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    • 제51권1호
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    • pp.57-62
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    • 2024
  • Objective: The purpose of this study was to use a mouse model to investigate the blastocyst formation rate in vitrified-warmed embryos derived from vitrified-warmed oocytes. Methods: Metaphase II oocytes obtained from BDF1 mice were vitrified and warmed, followed by fertilization with epididymal sperm. On day 3, a total of 176 embryos, at either the eight-cell or the morula stage, were vitrified-warmed (representing group 1). For group 2, 155 embryos at the same developmental stages were not vitrified, but rather were directly cultured until day 5. Finally, group 3 included day-5 blastocysts derived from fresh oocytes, which served as fresh controls. The primary outcome measured was the rate of blastocyst formation per day-3 embryo at the eight-cell or morula stage. Results: The rates of blastocyst formation per day-3 embryo were comparable between groups 1 and 2, at 64.5% and 69.7%, respectively (p>0.05). The formation rates of good-quality blastocysts (expanded, hatching, or hatched) were also similar for groups 1 and 2, at 35.5% and 43.2%, respectively (p>0.05). For the fresh oocytes (group 3), the blastocyst formation rate was 75.5%, which was similar to groups 1 and 2. However, the rate of good-quality blastocyst formation in group 3 was 57.3%, significantly exceeding those of group 1 (p=0.001) and group 2 (p=0.023). Conclusion: Regarding developmental potential to the blastocyst stage, vitrified-warmed day-3 embryos originating from vitrified-warmed oocytes demonstrated comparable results to non-vitrified embryos from similar oocytes. These findings indicate that day-3 embryos derived from vitrified-warmed oocytes can be effectively cryopreserved without incurring cellular damage.

Effect of oocyte chromatin status in porcine follicles on the embryo development in vitro

  • Lee, Joo Bin;Lee, Min Gu;Lin, Tao;Shin, Hyeon Yeong;Lee, Jae Eun;Kang, Jung Won;Jin, Dong-Il
    • Asian-Australasian Journal of Animal Sciences
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    • 제32권7호
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    • pp.956-965
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    • 2019
  • Objective: The main goal of this study was to provide a morphological indicator that could be used to select high-quality oocytes of appropriate meiotic and developmental capabilities in pig. The higher quality of immature oocytes, the higher success rates of in vitro maturation (IVM) and in vitro fertilization (IVF). Thus, prior to the IVM culture, it is important to characterize oocytes morphologically and biochemically in order to assess their quality. Two of the largest indicators of oocyte quality are the presence of cumulus cells and status of chromatin. To investigate the effects of porcine oocyte chromatin configurations on the developmental capacity of blastocysts, we assessed oocyte chromatin status according to follicle size and measured the developmental potency of blastocysts. Methods: To sort by follicle size, we divided the oocytes into three groups (less than 1 mm, 1 to 3 mm, and more than 3 mm in diameter). To assess chromatin configuration, the oocytes were assessed for their stages (surrounded nucleolus [SN] germinal vesicle [GV], non-surrounded nucleolus [NSN] GV, GV breakdown, metaphase I [MI], pro-metaphase II [proMII], and metaphase II [MII]) at different maturation times (22, 44, and 66 h). To assess the development rate, oocytes of each follicle size were subjected to parthenogenetic activation for further development. Finally, GV oocytes were grouped by their chromatin configuration (SN, SN/NSN, and NSN) and their global transcriptional levels were measured. Results: SN GV oocytes were more suitable for IVF than NSN GV oocytes. Moreover, oocytes collected from the larger follicles had a greater distribution of SN GV oocytes and a higher developmental capacity during IVM, reaching MII more quickly and developing more often to blastocysts. Conclusion: Porcine oocytes with high-level meiotic and developmental capacity were identified by analyzing the relationship between follicle size and chromatin configuration. The porcine oocytes from large follicles had a significantly higher SN status in which the transcription level was low and could be better in the degree of meiotic progression and developmental capacity.

아버지의 미취학자녀 돌봄시간 변화 추이 분석(2004-2019) (Trend in Paternal Childcare Time for Preschool Children in Korea from 2004 to 2019)

  • 이정은;서지원
    • 가족자원경영과 정책
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    • 제25권3호
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    • pp.103-120
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    • 2021
  • 본 연구는 맞벌이와 외벌이 아버지의 미취학자녀 돌봄시간의 변화를 파악하고 아버지의 자녀돌봄 활동의 특성 변화를 규명하기 위하여 돌봄의 하위활동영역을 각각 필수돌봄, 발달돌봄, 기타돌봄으로 분류하였다. 이를 위해 통계청 생활시간조사자료 중 2004, 2009, 2014, 2019년의 4개 연도 15년간의 자료를 사용하여 평일(근무일)의 아버지 자녀돌봄시간과 참여율을 분석하였다, 본 연구의 주요결과는 다음과 같다. 첫째, 아버지가 미취학자녀를 돌보는 시간은 증가 추이를 나타냈다. 특히, 맞벌이 아버지는 2004년과 비교하여 2019년에 자녀돌봄시간이 24분 증가하여 15년 동안 2배 가량의 증가를 보였다. 둘째, 아버지의 돌봄유형을 분석한 결과, 2004년에는 생존과 직결되지 않고 간헐적 참여가 가능한 발달돌봄 참여율이 자녀의 생존과 건강유지를 위한 필수돌봄에 참여율보다 적었으나, 2019년에는 외벌이 아버지는 필수돌봄과 발달돌봄 참여율이 유사하였고, 맞벌이 아버지는 필수돌봄 참여율이 더 높게 나타났다. 셋째, 아버지의 자녀돌봄시간에 영향을 미치는 변수에는 아버지의 연령, 교육수준, 성평등의식, 시장노동시간, 출퇴근시간 등이 통계적으로 유의하였다. 특히, 시장노동시간은 모든 조사시기에 영향을 미치는 변수였으며, 성평등의식은 외벌이 아버지 집단에서 최근까지 영향을 미치는 변수로 나타났다. 이상의 연구결과는 맞벌이 아버지와 외벌이 아버지 모두 자녀돌봄시간이 점차 증가하는 추세를 보였으며, 특히 발달돌봄뿐아니라 필수돌봄에 사용하는 시간이 늘어났다는 점에서 부모의 공동양육 책임이 실현되는 방향으로의 긍정적 변화를 시사한다. 또한, 이는 지난 15년간의 아버지 자녀돌봄 행태의 변화에 대한 이해를 위한 기초자료로 사용될 수 있으며, 향후 가족친화 정책의 구체적 과제를 제안하고 지역사회 등에서 아버지 대상 자녀돌봄 프로그램을 개발하는 근거자료로 활용될 것으로 기대된다.