• 한국수정란이식학회지
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• 제15권3호
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• pp.203-210
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• 2000

The objective of this study was to identify a follicular fluid ingredient inhibiting the cumulus oocyte complex (COC) expansion. Thus, follicular fluid or liquid chromatographic fractions of follicular fluid was supplemented in COC culture medium. And COCs were incubated for 48 hours to investigate about cumulus expansion and also the first polar body extrusion. The results obtained were as follows; 1. The fluid of medium follicle significantly inhibited the COC expansion. 2. The fluid of large follicle inhibited the COC expansion. 3. Follicular fluid showed six major fractions at retention volumes (RVs) 1.83, 1.91, 2.15, 2.34, 2.53 and 2.74 ml after separation with Superose 12 column. Of the major fractions, fractions RV2.15, RV2.34, RV2.53 and RV2.74 inhibited both COC expansion and polar body extrusion. Especially, fractions of RV2.15 and RV2.53 significantly inhibited COC expansion, oocyte denudation and polar body extrusion. In conclusion, porcine follicular fluid contained a COC expansion inhibiting ingredient (CEI) that may be contained largely in fractions RV2.15 and RV2.53. And CEI may inhibit oocyte maturation by inhibition of oocyte denudation and extrusion of the first polar body.

• 한국수정란이식학회지
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• 제5권1호
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• pp.21-27
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• 1990

The technique for maturation of follicular oocyte has been devised to provide such a low cost and in ptentifut number supply of bovine embryo. Some of problems concerning production of bovine embtyo in vitro were discussed in this paper. Bovine follicular oocytes cultured in vitro achieved normal fertilization but cleavage rates to blastocyst were low compared to the oocyte matured in vivo. It has been concluded that a deficient cytoplasmic maturation occurs in the oocytes matured in vitro. These results indicate that the studies for maturation of bovine follicular oocytes in vitro need improvement of culture conditions and to define the characteristics that might be indicative of healthy oocyte.

• Clinical and Experimental Reproductive Medicine
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• 제12권2호
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• pp.65-69
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• 1985

1) Rabbit follicular oocytes from preovulatory follicles were cultured for 12 hr in vitro and fertilized in vivo by transferring the oocytes to the first foster-mother. 2) Two youngs were bron from transferred embryos from the first foster-mother to the second foster-mother. This demonstrates that in vitro cultured follicular oocytes are normal and they can develop into normal young born when transferred to the foster-mother. 3) A simple chemically defined culture medium, salt sol. with glutamine (2mM), which was developed by Bae and Foote(1975) proves fully good enough for rabbit follicular oocyte culture. We call this B-F medium. 4) Twelve hours culture in vitro of the rabbit follicular oocyte may be a proper culture time for further development.

• 한국어류학회지
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• 제19권1호
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• pp.16-23
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• 2007

한국산 버들치속 (Rhynchocypris) 어류인 버들치 (Rhynchocypris oxycephalus)와 금강모치 (Rhynchocypris kumgangensis) 난모세포의 난막구조에 대해 광학현미경과 전자현미경으로 조사하였다. 두 종에 있어서 난형성과정은 비슷했으나 난모세포를 둘러싸는 여포세포층(follicular layer)에 있어서는 차이를 보였다. 버들치는 난황포(yolk vesicle)시기에 있어 여포세포층은 안쪽에 입방형 또는 둥근모양의 세포층(inner follicular layer)이 난막위에 형성되고 그 바깥쪽으로 편평세포층(outer follicular layer)의 2층으로 이루어져 있었다. 난모세포의 발생이 진행됨에 따라 inner follicular layer의 입방형세포는 원주형세포(columnar cell)로 바뀌게 된다. 난황구(yolk granule)시기에 원주형세포는 세포질에 부착물질인 mucin을 분비해서 난세포 전체를 둘러싸게 된다. 반면에 금강모치의 경우 버들치와 마찬가지로 난황포시기에 안층의 입방형 또는 둥근모양의 세포층과 바깥층의 편평세포층을 가지게 되지만 안층의 세포는 더 이상 변화를 보이지 않았으며, 부착물질 또한 형성되지 않았다. 이처럼 한국산 버들치속에 있어 난막의 구조적 차이는 두 종간에 뚜렷한 분류형질로도 이용될 수 있을 뿐 아니라 그들의 서식처 및 산란습성과도 연관이 있는 것으로 생각된다.

• Animal cells and systems
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• 제5권1호
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• pp.45-50
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• 2001

Rana ovarian follicles consist of oocyte, vitelline envelope, granulosa cells, and theca/epithelial layer. Using scanning electron microscopy, the surface structure of each follicular component was investigated. Changes in oocyte surface during oocyte maturation were also examined. Theca/epithelial layer was almost transparent and some blood vessels and granulosa cells were observed underneath in intact follicle. The number of granulosa cells was estimated to be 6700-7200 per oocyte. The granulosa cells partially overlapped each other and their microvilli penetrated the vitelline membrane via holes present in the vitelline envelope and seemed to be linked to oocyte microvilli. After removal of the vitelline envelope by microforcep, oocyte microvilli were observed on the surface of the devitellined oocyte. The oocyte microvilli formed partial clusters on the surface of white spot area which appears iust before germinal vesicle breakdown (GVBD), whereas they were evenly distributed in other areas. The microvilli became shorter and less dense with oocyte maturation. The lengths of oocyte microvilli in the immature and mature oocyte were 1.5 $\mu$m and 0.6 $\mu$m, respectively. The present study suggests a fundamental structural change occurring on the oocyte surface during maturation.

• Asian-Australasian Journal of Animal Sciences
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• 제18권11호
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• pp.1557-1563
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• 2005

Ovarian follicular fluid contains both stimulatory and inhibitory agents that influence the growth and maturation of oocyte. In the present study, an attempt was made to isolate and study the biological properties of ovarian follicular fluid peptide(s) in buffaloes. Bubaline ovarian follicular was made steroid- and cell-free. A protein fraction was obtained by saturation (30-35% level) of the follicular fluid with ammonium sulfate. The protein fraction was purified with Sephadex-G 50 gel filtration chromatography and a single peak was obtained in the eluant volume, which was lyophilized. SDS-PAGE of the lyophilized fraction revealed a single band and the molecular weight of the peptide was 26.6 kDa. The peptide stimulated the cumulus cell expansion and in vitro maturation rate of oocytes in buffaloes in a dose dependent manner when it was incorporated at different dose levels (0, 10, 25, 50, 100 and 1,000 ng $ml^{-1}$ of maturation medium). The basic culture medium consisted of TCM 199 with Bovine serum albumin (0.3%). The in vitro maturation rates were comparable to those obtained with a positive control medium (TCM 199+20 ng EGF $ml^{-1}$+steer serum (20%)). Further purification and biological assays may throw more light on the nature and functions of this peptide.

• Animal cells and systems
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• 제2권4호
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• pp.501-506
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• 1998

In the goby Micropercops swinhonis, the development of its egg's enveloping layers could be divided into 4 stages. In the earliest developmental period, stage I, there is a simple oocyte surrounded by a layer of squamous follicular cells. Stage II corresponds to the yolk vesicle stage of vitellogenesis. Here the initial follicular layer has become bilaminar with the retention of its outer squamous cell layer and the acquisition of an inner cuboidal cell layer just over the zona radiata. The number and size of the cuboidal cells increases throughout this stage. Stage III corresponds to the yolk granule stage of true vitellogenesis. Here the cuboidal cells begin to be replaced by columnar cells. As the oocyte grows, the columnar cells increase in size. The columnar cells produce cytoplasmic neutral mucins and by the end of this stage their cytoplasm has been filled with this mucin. In stage IV a single layer of squamous cells still remained as the outer follicular layer of the oocyte. The secretory activity of the inner follicular layers' columnar cells has ceased and they had lost their cell wall integrity and ended as a series of bullet-shaped, neutral mucin deposits.

• 한국수정란이식학회지
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• 제32권3호
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• pp.105-109
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• 2017

Autophagy is an intracellular degradation and recycling system. Oocyte maturation is dynamic process, in which various proteins should be synthesized and degraded. In our previous study, we reported the loci of autophagosome and dynamics of autophagic activity in porcine oocytes during in vitro maturation. In this study, we verified loci of autophagosome in porcine follicular cumulus-oocyte complex by detection of microtubule-associated protein 1A/1B-light chain 3 (LC3) which is the reliable marker of autophagosome. Porcine ovary including various sizes of follicles was fixed within 1 hour after collection from slaughterhouse. After fixation, immunohistochemistry was conducted on sliced ovary tissue containing various sizes of follicles by using LC3 antibody. As a result, LC3 signal was clearly detected in both cumulus and oocytes of various sizes of follicles. We also found ring shaped signal which represent autophagosome near oocyte membrane. Most of the signals in oocytes were localized nearby cellular membrane while evenly dispersed in cumulus cells. Therefore, this result suggests that autophagy occurs in porcine COCs (cumulus-oocyte complexes) at follicular stage.

• 한국가축번식학회지
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• 제14권1호
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• pp.84-91
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• 1990

본 실험은 돼지난포란의 체외성숙과 체외수정 효과를 높일 수 있는 방법을 찾기 위하여 시도되었으며 직경 1~2mm와 3~7mm 난포로부터 채란된 난자를 mKRB(-BSA)에 돼지발정혈청(ESS), FCS 또는 투석돼지난포액(DFF)을 첨가한 성숙배양에서 24~48시간, 37$^{\circ}C$에서 배양하였다. 성숙된 난포란은 정소상체 정자와 24시간 배양 후 전핵행성 여부를 조사하였다. 36~48시간 배양에서 50~60%의 난자가 metaphase II에 도달되었고 난포 크기(1~2mm와 3~7mm)간에 체외성숙율의 차이는 없었으나 3~7mm 난포란에서 성숙분열이 다소 빨랐다. 체외성숙배양액에 5% ESS, 15% FCS 및 DFF 첨가시 대조구보다 다소 성숙율이 높았다. 체외수정율(전핵형성)은 5% ESS와 15% FCS 첨가 성숙시킨 난포란과 체내 수정능획득 정자와의 수정에서 각각 높은 경향이 있었다. 따라서 돼지난포란의 체외성숙과 수정에 ESS, FCS 및 투석난포액이 유효한 요인이 됨을 알 수 있다.

• Clinical and Experimental Reproductive Medicine
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• 제41권1호
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• pp.21-28
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• 2014

Objective: To investigate the association of individual follicular fluid (FF) leptin and adiponectin levels with the quality of the corresponding oocyte and embryo. Methods: We prospectively enrolled 67 women who underwent controlled ovarian hyperstimulation with 89 FF samples. FF and the corresponding oocyte was obtained from a single dominant preovulatory follicle at the time of oocyte retrieval. Concentrations of leptin and adiponectin were measured by enzyme-linked immunosorbent assay in an individual follicle. The oocyte quality, fertilization rate, and corresponding embryo development were assessed. Results: The FF level of leptin was significantly associated with body mass index (r=0.334, p<0.01). The FF adiponectin level was significantly higher in the normal fertilization group than the abnormal fertilization group (p=0.009) in the non-obese women. A lower FF leptin level was associated with a trend toward mature oocytes, normal fertilization, and good embryo quality, although these relationships were not statistically significant. The leptin:adiponectin ratio of FF did not differ significantly according to oocyte and embryo quality. The quality of the oocyte and embryo was not associated with the FF leptin level tertile. However, the normal fertilization rate was positively associated with FF adiponectin level tertile. There was a trend towards improved oocytes and normal fertilization rates with the lowest tertile of the FF leptin:adiponectin ratio, but this difference was not statistically significant. Conclusion: Our results suggest that a high FF adiponectin concentration could be a predictor of normal fertilization. However, the FF leptin concentration and leptin:adiponectin ratio is not significantly related to oocyte maturity and corresponding embryo development.