• Title, Summary, Keyword: Follicular Fluid

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Studies on in vitro Maturation and Fertilization of Porcine Follicular Oocytes (돼지 난포란의 체외성숙 및 수정에 관한 연구)

  • 김상근;이만휘;이명헌;신용호
    • Korean Journal of Animal Reproduction
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    • v.14 no.1
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    • pp.23-30
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    • 1990
  • These studies were carried out to investigate the effects of fetal calf serum(FCS), estrous porcine serum(EPS), porcine follicular fluid(PFF), hormone and matured cumulus cell(MCC) on in vitro maturation and fertilization of porcine follicular oocytes. The ovaries and testes were obtianed from slaughtered Landrace sow and boars, respectively. The follicular oocytes surrounded with cumulus cells were recovered by aspirating follicular fluid from the visible follicles of diameter 3~5 mm and the semen were prepared from boar's epididymal cauda. The follicular oocytes were cultured in TCM-199 medium containing hormones, FCS, EPS, PFF and MCC for 48hrs. in a incubator with 5% CO2 in air at 36$^{\circ}C$ and then matured oocytes were again cultured for 18~20 hrs. with $1.5\times$106/ml motile capacitated sperm in the modified Tyroide solution containing 100$\mu\textrm{g}$/ml of heparin. The results obtained in these experiments were summarized as follows : 1. The maturation and fertilization rate of the follicular oocytes, cultured in the TCM-199 medium supplemented with 10% FCS and PMSG+HCG were 55.6~64.5% and 33.3~37.1%, respectively. 2. The maturation and fertilization rate of the follicular oocytes cultured in the TCM-199 medium supplemented with 20% EPS and PMSG+HCG were 50.0~55.0% and 30.3~33.3%, respectively. 3. The maturation rate(59.0~64.2%) and fertilization rate(34.8~39.3%) of follicular oocytes cultured in TCM-199 medium supplemented 20% FCS and 50% PFF were higher than those of follicular oocytes cultured in TCM-199 medium supplemented with 5%, 10% and 15% FCS and 10% and 50% PFF. 4. The maturation rate(60.0%) and fertilization rate(40.0%) of follicular oocytes cultured in TCM-199 medium supplemented with 20% FCS and granulosa cell (1$\times$106/ml) were significantly higher than those of fiollicular oocytes cultured in TCM-199 medium supplemented with 5%, 10% and 15% FCS and granulosa cell.

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Identification of Proteins in Human Follicular Fluid by Proteomic Profiling

  • Sim, Young-Jin;Lee, Mi-Young
    • Molecular & Cellular Toxicology
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    • v.4 no.3
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    • pp.253-259
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    • 2008
  • Human follicular fluid (HFF) is the in vivo microenvironment for oocyte maturation and includes a variety of proteins that could be involved in oocyte development and fertilization. We therefore used a proteomic approach to identify new HFF proteins. HFF from mature human follicles was obtained from five women following oocyte collection for in vitro fertilization (IVF). Ethanol-precipitated HFF run on two-dimensional gel electrophoresis (2DE) produced approximately 250 Coomassie brilliant blue-stained spots, 64 of which were identified using matrix-assisted laser desorption/ionization-mass spectrometry (MALDIMS). In this study, several proteins including complement factor H, inter-${\alpha}$ (globulin) inhibitor H4, inter-${\alpha}$-trypsin inhibitor heavy chain H4 precursor, human zinc-${\alpha}$-2-glycoprotein chain B, PRO2619, PRO02044, and complex-forming glycoprotein HC were new proteins that have not been previously reported in HFF using proteomic methods. Additionally, we identified alloalbumin venezia for the first time from trichloroacetic acid (TCA)-precipitated HFF. These HFF proteins could serve as new biomarkers for important human reproductive processes.

Changes in Number of Granulosa Cells, Follicular Fluid Levels and Diameter of Oocytes during Folliculogenesis in Pre-pubertal Gilts at Marketing Weight

  • Chiou, C.M.;Yang, T.S.;Yeh, S.P.;Tsai, M.Z.;Cheng, S.P.;Huang, M.C.
    • Asian-Australasian Journal of Animal Sciences
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    • v.17 no.12
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    • pp.1647-1651
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    • 2004
  • The follicles (1.8 to 7.8 mm in diameter) were recovered from the ovaries in marketed pigs and the number of granulosa cells, the diameter of oocytes obtained from different development stages of the follicles and follicular fluid levels were determined. Correlations between size measurements and cell counts as well as the diameter of antral follicles and oocytes were also investigated. The results indicated that, while expanding in size, follicle numbers decreased with a greater atretic proportion. Granulosa cells increased in numbers continuously and remained unchanged beyond the size of 200 ${mm}^3$ in non-atretic follicles, whereas a sudden drop of granulosa counts was observed in atretic follicles. Follicular fluid, on the other hand, linearly increased its volume with follicle size and differed little between those of non-atretic and atretic follicles. Diameters of oocytes in non-atretic follicles increased to its maximum when follicles expanded to 150 ${mm}^3$ and maintained its size during later follicular expansion. It is concluded that, for in vitro culture, the optimal size of porcine follicle should be between 150 to 180 ${mm}^3$if they are collected from pre-pubertal gilts of marketing size slaughtered in an abattoir.

Effects of Fetal Calf Serum and Porcine Follicular Fluid Fractionated by Gel Filtration on in vitro Maturation of Porcine Follicular Oocytes (Gel Filtration에 의해 분획된 소 태아혈청과 돼지난포액이 돼지난포란의 체외성숙에 미치는 효과)

  • 가학현;정구민;한정호;임경순
    • Korean Journal of Animal Reproduction
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    • v.19 no.4
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    • pp.251-258
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    • 1996
  • These studies were carried out to investigate the effect of gonadotropins (GTH), fetal calf serum (FCS), porcine follicular fluid (pFF) and FCS and pFF fractions obtained by the gel filtration on in vitro maturation of porcine follicular fluid. When the oocytes were cultured in TCM-199, the maturation rate was higher in pFF than in FCS in both with or without GTH and in pFF the maturation rate was higher in with GTH than in without GTH. In case of without GTH, pFF increased maturation rates in TCM-199, but not in Whitten's medium (WM). When the oocytes were cultured in WM supplemented with FCS fractions, the maturation rate(51.6%) of oocytes was significantly (P<0.05) higher in fraction B (about 30∼70 kDa) than in control, FCS and other fractions. When oocytes were cultured in WM supplemented with pFF fractions, fractions B (about 30∼70 kDa) and D (about 1∼10 kDa) were significantly (P<0.05) higher than in control, pFF and other fractions. In conclusiion, the addition of gonadotropins into the maturation media was effective for oocyte maturation. The addition of pFF was more effective than addition of FCS for maturation of porcine oocytes in vitro. And fraction B from FCS and fractions B and D from pFF was effective for oocyte maturation.

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Change of Protein Patterns in Uterine Fluid during Estrus Cycle in Pigs (돼지에서 발정 주기 동안 자궁액 내의 단백질 패턴의 변화)

  • Lee, Yeon-Ju;Song, Eun-Ji;Lee, Sang-Hee;Cheong, Hee-Tae;Yang, Boo-Keun;Park, Choon-Keun
    • Reproductive and Developmental Biology
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    • v.37 no.3
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    • pp.103-108
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    • 2013
  • An uterus is female reproductive tract organ that affected estrus cycle. During a various changes occur at uterus in estrus cycle, one of them is body fluids secretion be called uterine fluid. Therefore, the objective of this study was to investigate the changes of protein patterns using two-dimensional gel electrophoresis in uterus fluids during the follicular and luteal phases in estrus cycle of pigs. In changes of protein spots were confirmed during the follicular and luteal phases. The 136 spots were expressed in follicular phase, the 57 spots of them showed reproducibility. On the other hand, the 140 spots were expressed in luteal phase, the 73 spots of them showed reproducibility. Also, spots expressed in follicular phase were number 69 and 94 spots and spots expressed in luteal phase only were number 156, 157, 184~187, 190 and 191 spots. The spots which of higher expression levels in the luteal phase than in follicular phase were number 76 and 79 spots. In conclusion, the spots expressed in follicular and luteal phases were confirmed with difference levels and these differences are function of RNA resolving, protein synthesis and cytoskeletal architecture.

Study on functional states and differential diagnosis of ovarian cysts in slaughtered Korean native cows (도축한우에서 난소낭종의 감별진단 및 기능성에 관한 연구)

  • 이청산;류대열;윤호규;송종한;이종인;서국현;김일화;강현구
    • Korean Journal of Veterinary Service
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    • v.27 no.1
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    • pp.41-52
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    • 2004
  • To establish the differential diagnosis and functional status in ovarian cystic cows, progesterone(P$_4$) and estrogen(E$_2$) level of cystic follicular fluid, ultrasonography for measuring the cystic diameter and thickness of cystic wall, and histological findings were investigated in cystic ovaries from slaughtered Korean native cows. Ovarian follicles were classified as systic if the diameter was greater than 25 mm by ultrasonography. Ovarian cysts < 3 mm of cystic wall thickness, < 10 ng/ml P$_4$ concentration and >10 ng/ml E$_2$ concentration were classified follicular cyst, ovarian cysts 3 mm of cystic wall thickness, 10 ng/ml P$_4$ concentration and <10 ng/ml E$_2$ concentration were classified luteal cyst, and ovarian cysts 3 mm of cystic wall thickness, < 10 ng/ml P$_4$ concentration and <10 ng/ml E$_2$ concentration were classified non-functional ovarian cyst, respectively. Also ovarian cysts were classified 8 types by anatomical and hisctological findings. Ovarian cysts with corpus luteum were 3 of 73 cows and ovarian cysts without corpus luteum were 70 cows. The incidence rates of 8 various types of ovarian cysts were as follows; 2Aa 56.2%, 2Ba 20.5% and 2Ab 15.1%, respectively. The incidence rates of ovarian cysts without corpus luteum were follicular cyst 76.7% and luteal cyst 19.2%. The thickness of cystic wall were lAb 3.9 mm, 2Ab 3.3 mm and 2Bb 3.2 mm, and the cystic fluid P$_4$ concentrations were above 10.0 ng/ml in lAb, 2Ab and 2Bb, respectively. There was significantly correlations between the thickness of cystic wall and cystic fluid P$_4$ concentration in ovarian cysts(p<0.05). The ovarian cyst was classified follicular cysts, luteal cyst and non-functional ovarian cyst by hormone analysis. The luteal cyst was accuratly dignosed by cystic wall thickness. But follicular cysts was misdiagnosed 13 cows of 56 cystic cows. The 13 cystic cows was determined as had non-fuctional ovarian cysts. The cystic fluid P$_4$ concentration was 3.3 ng/ml in follicular ovarian cysts and 30.1 ng/ml luteinized ovarian cysts. There was significantly positive correlations between thickness of cystic wall and serum P$_4$ concentration in follicular(r$^2$ =0.59, p<0.001) and luteal cysts(r$^2$=0.65, p<0.001). These results indicated that ovarian cysts had various stages of degeneration and luteal cyst was accuratly diagnosed measurement of cystic wall thickness by ultrasonography, but follicular cysts was not diagnosed only cystic diameter and cystic wall thickness. In conclusion, it is suggest that ovarian cysts was diagnosed by combination of clinical sign and anatomical cystic features.

Studies on the Concentrations of Sex Hormone in the Blood Plasma and Antrum Fluid of Follicular and Lutein Cystic Ovaries of Holstein Cows (난포낭종 및 황체난종우의 혈장과 낭종내강액내 성호르몬의 농도에 관한 연구)

  • 김상근;임영재
    • Korean Journal of Animal Reproduction
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    • v.13 no.2
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    • pp.85-92
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    • 1989
  • The study was carried out to find out the changes of the sex hormone concentrations in the blood plasma and antrum of follicular and lutein cystic ovaries of Holstein cows. The progesterone, estradiol-17$\beta$, testosterone, FSH and LH from samples of the blood plasma and antrum of cystic ovaries of cows assayed by radioimmunoassay method. The results of this study were summarized as follows : 1. The concentrations fo progesterone and estradiol-17$\beta$ in the blood plasmaat estrous and luteal phase were 0.95$\pm$0.18ng/ml, 11.45$\pm$3.12pg/ml and 4.25$\pm$0.27ng/ml, 6.27$\pm$0.82pg/ml respectively. The concentrations of progesterone and estradiol-17$\beta$ in the antrum fluid of follicles at estrous and luteal phase were 24.8$\pm$4.12ng/ml, 54.3$\pm$7.25pg/ml and 21.7$\pm$3.79ng/ml, 14.3$\pm$2.72pg/ml respectively, and showed significant changes among the estrous and luteal phase and blood plasma and antrum fluid of follicles. 2. The concentrations of progesterone, estradiol-17$\beta$, testosterone and LH in the blood plasma of follicular cystic ovareis of cows were 0.85$\pm$0.25ng/ml, 9.23$\pm$2.72pg/ml, 17.12$\pm$3.26pg/ml and 3.78$\pm$1.02mIU/ml respectively. And the concentrations of progesterone, estradiol-17$\beta$, testosterone and LH in the antrum fluid of follicular cystic ovareis of cows were 284$\pm$48.21ng/ml, 389$\pm$67.23ng/ml, 12.84$\pm$0.29ng/ml and 1.84$\pm$0.17mIU/ml respectively, and showed significant changes between in the blood plasma and antrum fluid of cystic ovaries. 3. The concentrations of progesterone, estradiol-17$\beta$, testosterone and LH in the blood plasma of lutein cystic ovaries of cows were 3.40$\pm$0.78ng/ml, 4.02$\pm$0.42pg/ml, 10.72$\pm$2.74pg/ml and 0.76$\pm$0.12mIU/ml respectively. And the concentrations of progesterone, estradiol-17$\beta$, testosterone and LH in the antrum fluid of lutein cystic ovareis of cows were 427$\pm$35.79ng/ml, 0.76$\pm$0.07ng/ml, 3.45$\pm$0.57ng/ml and 0.29$\pm$0.07mIU/ml respectively, and showed significant changes between the blood plasma and antrum fluid of cystic ovaries. 4. Accordingly, the diagnosis of follicular and lutein cystic ovareis of cows from progesterone, estradiol-17$\beta$ and LH levels in the blood plasma and antrum of cystic ovaries of cows is thought to be possible a diagnostic means.

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Follicular fluid cerebellin and betatrophin regulate the metabolic functions of growing follicles in polycystic ovary syndrome

  • Ersahin, Aynur Adeviye;Acet, Mustafa;Ersahin, Suat Suphan;Acet, Tuba;Yardim, Meltem;Kenanoglu, Omer;Aydin, Suleyman
    • Clinical and Experimental Reproductive Medicine
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    • v.44 no.1
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    • pp.33-39
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    • 2017
  • Objective: The aim of this study was to assess the changes of follicular fluid (FF) and serum levels of cerebellin precursor protein 1 (cbln1) and betatrophin in patients with polycystic ovary syndrome (PCOS) undergoing in vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI) with a gonadotropin-releasing hormone (GnRH) antagonist protocol. Methods: Twenty infertile women with PCOS and 20 control women diagnosed as poor responders undergoing ovarian stimulation with a GnRH antagonist were included. Blood samples were obtained during ovum pick-up. Follicular fluid from a dominant follicle was collected from the subjects. Using enzyme-linked immunosorbent assays, FF and serum levels of cbln1 and betatrophin were measured in both groups of participants. Metabolic and hormonal parameters were also determined and correlated with each other. Results: Both groups of women had similar serum and FF betatrophin levels ($55.0{\pm}8.9ng/mL$ vs. $53.1{\pm}10.3ng/mL$, p=0.11). The serum and FF betatrophin levels of poor responders were found to be similar ($49.9{\pm}5.9ng/mL$ vs. $48.9{\pm}10.7ng/mL$, p=0.22). Conversely, the FF cbln1 levels of PCOS women were found to be significantly higher than the serum cbln1 levels ($589.1{\pm}147.6ng/L$ vs. $531.7{\pm}74.3ng/L$, p<0.02). The FF cbln1 levels of control participants without PCOS were significantly higher than their serum cbln1 levels ($599.3{\pm}211.5ng/L$ vs. $525.3{\pm}87.0ng/L$, p=0.01). Positive correlations were detected among body mass index, insulin resistance, serum insulin, total testosterone, and betatrophin levels in the PCOS group. Conclusion: Follicular fluid betatrophin and cbln1 concentrations may play a pivotal role on follicular growth in PCOS subjects undergoing IVF/ICSI with an antagonist protocol.