• Title/Summary/Keyword: Oocyte size

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Correlation of Oocyte Quality with Fertilization and Embryonic Development in Intracytoplasmic Sperm Injection Treatment (세포질내 정자주입술 시행시 난자의 상태에 따른 수정률과 배아의 발생양상)

  • 임천규;전진현;송상진;김정욱;강인수
    • Development and Reproduction
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    • v.2 no.2
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    • pp.213-221
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    • 1998
  • Intracytoplasmic sperm injection (ICSI) has been widely used to treat couples with infertility due to severely impaired sperm charateristics and for whom conventional in-vitro fertilization (IVF) had failed. The extent to which the morphology of the oocyte at the light microscopy level is related to the results of ICSI vis controversial. In this study, oocytes from 44 patients were reviewed. The ICSI procedure was recorded through CCD camera. The oocytes were divided into five groups according to the presence of cytoplasmic inclusions, the width of perivitelline space (PVS), the presence of cell debris in PVS, the status of first polar body and the flexibility of oolemma. The results showed that the fertilization rate and embryonic development were not associated with the morphological criteria of oocyte. The degeneraton rate of oocytes after ICSI was significantly higher (P<0.001) in the oocytes whose membranes were broken at the moment of insertion (17.7%) than the oocytes whose membranes were broken by aspiration of cytoplasm (1.6%). More oocytes with cytoplasmic inclusions (48.4% vs. 25.1%, p<0.001), wide PVS (35.2% vs. 19.0%, p<0.001), or cell debris in PVS (53.3% vs. 38.4%, p<0.05) were observed in patients with female factor infertility compared to patients with male factor infertility. These results .suggest that the fertilization rate and embryonic development after ICSI are not correlated with oocyte morphology based on the presence of cytoplasmic inclusions, size of PVS, the presence of cell debris in PVS and the status of polar body. And the degeneration rate of oocytes after ICSI was associated with the flexibility of oolemma.

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Pregnancy and Survival Rate of In Vitro Fertilized Bovine Embryos Frozen for Direct Transfer (직접이식을 위한 소 체외 수정란의 동결 융해후 생존성 및 수태율에 미치는 영향)

  • 오성종;양보석;이명식;백광수;성환후;정진관;임경순
    • Korean Journal of Animal Reproduction
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    • v.19 no.1
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    • pp.49-54
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    • 1995
  • This experiments were carried out to investigate the viabilities and the pregnancy rate of frozen-thawed IVF bovine embryos in various media, cryoprotectants and age of embryos produced. Hanwoo oocyte were collected in size of 2~7mm follicles, matured for 20~22hrs at 38.5$^{\circ}C$ in 5% CO2 incubator and then in vitro fertilized with Hanwoo semen. Blastocysts or more developed embryos at Day 7, 8 and 9 were frozen in 1.5 or 1.8M ethylene glycol. Viability of frozen thawed IVF embryos were identified the reformation of blastocoele after thawing and culture for 24~48 hours at 38.5$^{\circ}C$ in 5% CO2 incubator. Production rate of Hanwoo IVF embryos in TCM 199 and CR1aa ws 21.3%(39/183) and 28.1%(41/146), respectively. The viability of frozen thawed IVF embryos was higher rate in 1.8M ethylene glycol and Day 7 embryos than that in 1.5M and Day 8.53 cows out of 100 Hanwoo receipients transfered IVF embryos were pregnant and twin production rate was 26.3%.

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Seasonal Changes in Reproductive Condition of the Pacific Oysters, Crassostrea gigas (Thunberg) from Suspended Culture in Gosung Bay, Korea

  • Thao T. T. Ngo;Kang, Sang-Gyun;Park, Kwang-Sik
    • Korean Journal of Environmental Biology
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    • v.20 no.3
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    • pp.268-275
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    • 2002
  • Seasonal variation in reproductive condition of the Pacific oyster Crassostrea gigas was investigated from a suspended cultured oyster population in Gosung Bay, South Korea using histological techniques, Gametogenesis of oysters initiated in February when water temperature reached 11 to $13^\circ{C}$. Increase in oocyte size and the number resulting in follicle expansion was observed from March to May First spawning of oysters observed in mid Jun when the surface water temperature reached 22 to $25^\circ{C}$. Spawning activity of oysters extended from mid June to late September with two marked spawning peaks in June and August. Most oysters collected from October to December exhibited few residual eggs in packed follicles exhibiting a typical spent condition. No gametes were observed from December to February from oysters collected in the Bay. Gonadal development of oysters in the Bay seemed to follow a seasonal fluctuation in environmental conditions such as water temperature and food availability in the water column. Spawning of oysters in late June was in part associated with sudden drop in salinity due to vast amount of freshwater input in the Bay after the summer flooding. Sex ratio of oysters was 59.5% male and 39.8% female. Less than 1 percent (0.6%) of the oysters examined were hermaphrodite; few eggs were observed in testis.

In Vitro Growth and Development of Mouse Preantral Follicles

  • Kim, Dong-Hoon
    • Proceedings of the KSAR Conference
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    • 2000.10a
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    • pp.9-9
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    • 2000
  • The mammalian ovary has a large number of primordial and preantral follicles, which are a potential source of oocytes for the in vitro mass production of embryos. Several in vitro culture systems have been developed to support the growth and development of oocytes from mouse preantral follicles. Under the appropriate condition, meiotically incompetent oocytes from preantral follicles can grow to final size and complete nuclear maturation in vitro. Furthermore, the successful production of live young from in vitro grown and matured oocytes demonstrates that oocytes from preantral follicles are able to acquire full developmental capacity in vitro. However, the efficiency of in vitro production of embryos from mouse preantral follicles is still low. In farm animals as well as human, the growth of oocyte from preantral follicle to the meiotic competence stage has yet to be demonstrate. Therefore, further studies to improve the culture condition or to develope new culture system should be needed in the future. In addition, the visible progress in the establishment of the in vitro culture system for preantral follicles of farm animals and human could help to enlarge the populations of valuable agricultural, phamaceutical product-producing, and endangered animals, and to rescue the oocytes of women about to undergo clinical procedures that jeopardize oocytes.

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Organ Culture of Ovary Isolated from Juvenile Mice (약령 마우스에서 분리한 난소의 기관배양에 관한 연구)

  • 이현주;김지철;김기동;이상호;송해범
    • Journal of Embryo Transfer
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    • v.17 no.3
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    • pp.195-201
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    • 2002
  • This study was to assess the developmental capacity of oocytes matured in vitro after 20, 15, 10, 5 and 0 days of organ culture when ovaries were isolated from juvenile mice at 0-, 5-, 10-,15- and 20-day old, respectively, and to develop in vitro culture system that observed a view to morphology of ovaries and nucleus maturation of oocytes. The size of ovaries decreased 35.9%, 8.7%, 1.2% and 14.4% after 20, 15, 10, 5 days of organ culture when the ovaries were isolated from 0-, 5-, 10 and 15-day old mice, respectively. After organ culture, the recovery rates, diameters of oocytes and the number of oocytes progressed from GV to MII were increased as increasing age of mice.

Involvement of GTP-Binding Proteins in Stage-Specific Receptor-Mediated Endocytosis of Coelomic Fluid Proteins into Oocytes of Pseudopotamilla occelata (안점의 꽃갯지렁이 난포세포로 체강액 단백질의 단계특이적 유입을 위한 GTP-Binding Protein의 개입)

  • 남현정;강화선;이양림
    • The Korean Journal of Zoology
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    • v.39 no.3
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    • pp.292-298
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    • 1996
  • Receptor-mediated endocytosis of coelomic fluid proteins (CP), yolk precursor proteins, appears to be regulated by multiple GTP-binding proteins during oogenesis of a polychaete, Pseudopotamilla occelata. Transport of 125 I-CP into the oocytes of intermediate size class, at which CP is the most actively transported, is enhanced by GTP but inhibited by GTP analogues, either GTPrS or GTP$\beta$S. The effects of GTP and GTPrS on the transport were also confirmed by tracing internalization of gold-labeled CP with transmission electron microscope. Internalization of gold-labeled CP into the yolk granules was enhanced by GTP but inhibited by GTPrS.

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A Study on the Reproduction of the Sunrise Sculpin Pseudoblennius cottoides in Korea (한국산 가시망둑(Pseudoblennius cottoides)의 생식에 관한 연구)

  • Yoo, Dong Jae;Park, Jae Min;Lee, Sung Hun;Han, Kyeong Ho
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.50 no.1
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    • pp.48-54
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    • 2017
  • The ecological characteristics of Pseudoblennius cottoides were investigated off Dolsan Island, Yeosu, Korea, from June to December, 2001. Their habitats were the reef zone and surrounding embankment which was covered with seaweeds. Their range of total length (TL) was 6.10-8.40 cm (n=15), and the individuals (over 8.10 cm) of them accounted for in June. It was 8.60-15.7 cm (n=67), and the individuals 11.1-12.0 cm in TL accounted for in October, 2001. The female to male sex ratio was 1:0.48 (n=185). The gonads were in the shape of a rod and the ovaries were large in size. In June, the histology of the seminiferous tubules was observed. In August, the testicles appeared. In October, spermatocyte was observed in the testes, and ovaries were observed in relation to circumference popularity and oocyte.

In vitro maturation of porcine follicular oocytes (돼지 난포내 난모세포의 체외성숙에 관하여)

  • Park, Mi-hee;Lee, Hyo-jong
    • Korean Journal of Veterinary Research
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    • v.32 no.1
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    • pp.135-142
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    • 1992
  • This experiment was carried out to establish an effective technique of in vitro maturation of porcine follicular oocytes. Porcine ovaries were collected from an abbatoir and delivered to the laboratory in phosphate buffered saline in an hour. Immatured follicular oocytes were collected from the ovaries and divided into groups by the size of follicles and by the attachment of granulosa cells. The follicular oocytes were cultured in m-KRB solution supplemented with FCS(10%), follicular fluid(10%) or hormones of PMSG(10IU/ml), hCG(10IU/ml ) and $estradiol-17{\beta}(1{\mu}g/ml)$ for 48 hours at $39^{\circ}C$ under an atmosphere of 5% $CO_2$ in air. The results are as follows ; 1. The mean recoveration rate of follicular oocytes was 61.8%. 2. The maturation rate was significantly(p<0.05) higher when the oocytes were collected from large-sized follicles and under good state of granulosa cell attachment. 3. The maturation rate was significantly(p<0.01) promoted when the follicular oocytes were cultured in m-KRB solution supplemented with follicular fluid(74.8%) or hormones and fetal calf serum(70.6%).

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CHANGES OF THE CONCENTRATIONS OF STEROID HORMONES IN THE PORCINE FOLLICULAR FLUIDS ON ATRESIA (돼지여포의 퇴화과정 중 여포액 내 스테로이드 호르몬 양의 변화)

  • Lee, Young-Keun;Yoon, Yong-Dal
    • Clinical and Experimental Reproductive Medicine
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    • v.12 no.1
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    • pp.83-98
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    • 1985
  • In order to study the mechanism of follicular atresia, the follicles of the porcine ovary were isolated according to the presence or absence of the corpus luteum and their size, and then classified to the normal? or atretic?follicle on the morphological observation such as the transparency, the vascularization of follicle, the nuclear phase of oocyte, and the homogeneity of the granulosa cell layer. The viability of granulosa cells was examined. The concentrations of progesterone ($P_4$), testosterone (T), and estradiol-17 beta ($E_2$) in each follicular fluid were estimated by the radioimmunoassay. The viability of granulosa cells in the atretic follicle was much lower than that of the normal one. The concentration of each steroid hormone increased as the follicular size was increased, was not different in quantity between the normal- and the atretic follicle of which diameter was below 3mm, and were much higher in the atretic follicle than those in the normal one of which diameter was above 7mm. The ratio of the concentration of E2 to T in the large atretic follicle valued higher than that in the normal one, but smaller in the small and medium atretic follicle than that in the normal one. The present study suggests that the mechanism of atresia of the large follicle may be different from that of the small and the medium follicle and that the amount of steroid hormones regarded as the one of the criteria for the atretic follicles.

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A Comparative Study on the Parthenogenetic Development of Pig Oocytes Cultured in North Carolina State University-23 and Porcine Zygote Medium-3

  • Lee, Joo-Hyeong;Hyun, Sang-Hwan;Lee, Eun-Song
    • Journal of Embryo Transfer
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    • v.27 no.2
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    • pp.121-126
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    • 2012
  • The objective of this study was to examine the effect of in vitro culture media on embryonic development of in vitro-matured (IVM) oocytes after parthenogenetic activation (PA) in pigs. Immature pig oocytes were matured in TCM-199 supplemented with porcine follicular fluid, cysteine, pyruvate, EGF, insulin, and hormones for the first 22 h and then further cultured in hormone-free medium for an additional 22~26 h. IVM oocytes were activated by electric pulses and cultured in porcine zygote medium-3 (PZM-3) and North Carolina State University-23 supplemented with essential and non-essential amino acids (NCSU-23aa). These media were further modified by supplementing 2.77 mM myo-inositol, 0.34 mM trisodium citrate, and $10{\mu}M$ ${\beta}$-mercaptoethanol (designated as mPZM-3 and mNCSU-23aa, respectively). Culture of PA embryos in mPZM-3 significantly increased development to the blastocyst stage than culture in NCSU-23aa (36.2% vs. 24.8%, p<0.05). Modified PZM-3 showed a significantly higher blastocyst formation than NCSU-23aa in both groups of embryos that were activated at 44 h and 48 h of IVM (51.0% vs. 35.5% and 49.0% vs. 34.2% in oocytes activated at 44 h and 48 h of IVM, respectively). Irrespective of the follicle diameter where oocytes were collected, embryonic development to the blastocyst stage was increased (p<0.05) by the culture in mPZM-3 compared to culture in NCSU-23aa (25.9% vs. 34.2% and 32.9% vs. 44.8% in embryos derived from small and medium size follicles, respectively). Our results demonstrated that culture media had significant effect on preimplantation development PA embryos and that mPZM-3 was superior to mNCSU-23 in supporting development to the blastocyst stage in pigs. This beneficial effect of mPZM-3 on embryonic development was not impaired by other factors such as time of oocyte activation and origin of immature oocytes (small and medium size follicles).