• Title, Summary, Keyword: Follicular Fluid

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Study on Biochemical Constituents of Caprine Ovarian Follicular Fluid after Superovulation

  • Mishra, O.P.;Pandey, J.N.;Gawande, P.G.
    • Asian-Australasian Journal of Animal Sciences
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    • v.16 no.12
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    • pp.1711-1715
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    • 2003
  • The experiment was designed on 42 non pregnant Black Bengal goat. Out of which 18 were subjected to a superovulatory treatment comprising of eCG and hCG for embryo transfer study. The remaining 24 goats received no treatment and served as control for parameter studied as well as recipient for embryo transfer studies. Important biochemical constituents such as acid and alkaline phosphatase, total protein and cholesterol and inorganic phosphorus were estimated in the follicular fluid of control and treated group and the values were separately recorded for small medium and large size follicle. The results indicated a significant effect on acid phosphotase activity due to size of follicle. The value increased progressively from small to medium and from medium to large follicles. Alkaline phosphotase activity showed reverse trend. Alkaline phosphotase decreased progressively as size increased. The concentration of inorganic phosphorus did not reveal any significant difference between the control and treatment groups and also between the different size follicles. The concentration of protein decreased significantly from small to medium and from medium to large, although no difference was observed between the control and treatment groups. The concentration of Cholesterol in the follicular fluid indicated a significant increase from small to medium and to large follicle. Here also no difference was observed due to treatment. Similar in the composition of follicular fluid in the respect of above mentioned constituents indicated no of super ovulatory treatment on follicular fluid composition.

Effect of Follicular Fluid Proteins and Gonadotropins on Progesterone Secretion by Buffalo Granulosa Cells In vitro

  • Vinze, Mukesh;Sharma, M.K.;Singh, Dheer
    • Asian-Australasian Journal of Animal Sciences
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    • v.17 no.11
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    • pp.1496-1500
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    • 2004
  • In the mammalian ovary the follicular fluid contains proteins and peptides which play an important role in growth, development and maturation of oocytes. The gonadotropins and some other factors work synergistically and regulate ovarian functions. In the present study the effect of follicular fluid proteins (FFP) and gonadotropins on progesterone secretion by granulosa cells (GC) from buffalo ovary, was investigated during culture. The follicular fluid was collected from small (<5 mm), and medium (5-8 mm) follicles obtained from buffalo ovaries. The follicular fluid from medium follicles was fractionated with ammonium sulphate at 80% saturation. The precipitated protein fraction was further resolved in to minor (peaks I, III) and major (peak II) proteins using gel filtration (Sephadex G-200). The FFP from small follicles and major FFP (peak II) at a dose of 200 $\mu$g/well, significantly stimulated progesterone secretion by pooled GC (3${\times}10^{5}$ cells/2 ml medium/well). The minor FFP did not show any stimulatory effect. There was a significant increase in progesterone secretion by pooled GC in presence of FFP and LH (10 ng/well), however, FSH (20 ng/well) with FFP exhibited an inhibitory effect. The major FFP and gonadotropins were also studied for their effect on progesterone production by GC isolated from medium and large size follicles. The GC from medium follicles were more responsive to FSH and FFP whereas GC from large follicles exhibited enhanced progesterone secretion with LH and FFP. These results indicated that FFP have their own stimulatory effect and also act synergistically with gonadotropins. The significantly different response shown by GC, for steroid hormone secretion, is based on their stage of growth and differentiation. The purification and characterization of such steroidogenic proteins may help in elucidating their role in growth and differentiation of granulosa cells.

Effect of Maturation Medium and Porcine Follicular Fluid on In Vitro Maturation of Porcine Oocytes (배양액 및 난포액이 돼지 난포란의 체외성숙에 미치는 영향)

  • 박병권;이규승;박창식;서길웅
    • Korean Journal of Animal Reproduction
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    • v.20 no.3
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    • pp.289-297
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    • 1996
  • This study was conducted to investigate the effects of maturaton medium and porcine follicular fluid on in vitro maturation of porcine follicular oocytes. The results obtained are as follows; 1. When the oocytes were cultured for 42 hours, the maturation rate was significantly (P<0.05) higher in TCM-HEPES(75.5%) than Ham's F-10(60.9%) and mKRB(60.7%) medium. The optimal medium for the maturation of porcine oocytes in vitro was the TCM-HEPES medium. 2. When the oocytes were cultured for 42 hours in TCM-HEPES medium with 10, 20 and 30% of porcine follicular fluid(pFF), the maturation rates of porcine oocytes were 69.1, 65.6 and 63.3%, respectively. The maturation rate(51.4%) of oocytes cultured without pFF was significantly(P<0.05) lower than that of oocytes cultured with pFF. 3. The maturation rates of porcine oocytes cultured for 42 hours in TCM-HEPEs medium with 3 different porcine follicular fluid treatments were 68.6%(centrifused), 72.3%(filtered) and 73.1%(heat treated), respectively. The maturation rate(49.4%) of control group without pFF treatment was significantly(P<0.05) lower than that of oocytes cultured with pFF treatment.

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Effect of Follicular Fluid on Sperm Swim-up Separation with Sucrose Layer (난포액이 Sucrose 층을 이용한 정자의 Swim-up 분리에 미치는 효과)

  • 김경화;여영근;박영식
    • Journal of Embryo Transfer
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    • v.13 no.3
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    • pp.277-289
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    • 1998
  • To establish a system for sperm swim-up separation through sucrose layer, indiscreet sperm migration should sufficiently to block but movement of sperm shouldn't inhibit. Thus, the effects of sucrose levels in sucrose layer, incubation times and types of sucrose layer on sperm separation were examined. And the results obtained were as follows; 1. Layer of 10mM sucrose inhibited sperm swim-up migration through sucrose layer. 2. Incubation for 25 minutes without sucrose layer significantly increased sperm swim-up migration. However, incubation for 10 minutes to induce swim-up through sucrose layer significantly stimulated sperm migration and maintained sperm movement. 3. There was no significant difference between Type I and Type II in barrier effect of sucrose layer. However, sucrose layer of Type II with shorter distance of barrier was efficient for sampling. To elucidate a function of follicular fluid on sperm chemotaxis using in vitro system of sucrose layer of Type II and incubation for 10 minutes, the effects of dilution, heat treatment, and protein and lipid extracts of follicular fluid on sperm swim-up separation were examined. And the results obtained were as follows; 4. Follicular fluid stimulated sperm migration and movement, and significantly-attracted capacitated-sperm at 10% level. 5. Follicular fluid heated at 55$^{\circ}C$ for 30 minutes maintained the effect of follicular fluid stimulating sperm migration and movement. 6. Follicular protein stimulated sperm movement that was reduced by filtration of the protein. 7. Follicular lipid didn't significantly stimulate sperm migration and movement. 8. Both of follicular protein and lipid reduced the effect of follicular fluid stimulating sperm migration and movement. In conclusion, sucrose layer could be used for a barrier against indiscreet sperm migration by swim-up. And follicular fluid stimulated migration and movement of sperm and attracted capacitated-sperm through sucrose layer. Especially, heat-resistant protein of follicular fluid stimulated sperm migration.

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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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The Relationship Between the Pyknosis of Granulosa Cell and the Change of Steroid Concentration in Porcine Ovarian Follicles (돼지 난소내 과립세포의 염색질이상응축과 여포액내 스테로이드호르몬의 농도 변화와의 상관관계)

  • Lee, Chang-Joo;Yoon, Yong-Dal;Kim, Jong-Heup;Kim, Moon-Kyoo
    • Clinical and Experimental Reproductive Medicine
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    • v.16 no.1
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    • pp.35-40
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    • 1989
  • In order to study the follicular atretic mechanism in mammalian ovary, the relationship between pyknotic index (PI) of granulosa cells and the steroid concentrations in the follicular fluid of atretic follicle was investigated. Follicles were isolated from porcine ovary according to their sizes and the reproductive phases. Steroid concentrations were quantified by high-performance liquid chromatography. As PI increased, the concentration of progesterone was significantly increased(p<0.05), whereas testosterone and estradiol showed no significant changes in their concentration. As follicular size was increased, PI of follicular GC in the luteal phase was increased significantly(p<0.05) and the molar ratio of progesterone to testosterone was increased in the follicles of follicular phase. It can be concluded that progesterone accumulated in the follicular fluid as atresia of the follicles was progressed, and that PI of granulosa cells could be used as one of convenient and pratical criteria for the identification of follicular atresia.

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Effect of Rabbit Peritoneal Fluid(PF) on in vitro Maturation of Porcine Follicular Oocytes (가토복수가돼지 난포란의 체외성숙에 미치는 효과)

  • 정형민;박세필;오종훈;이훈택;정길생
    • Korean Journal of Animal Reproduction
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    • v.15 no.3
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    • pp.221-224
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    • 1991
  • This study was undertaken to evaluate the effect of rabbit peritioneal fluid(rPF) on in vitro maturtion of porcine follicular oocytes. From does 20h after hCG injection, rPF was aspirated aseptically at laparatomy, and then centrifuged, filtrated, and preincubated immediately for 12h. Porcine follicular oocytes isolated from ovaries of slaughtered animals were incubated in TCM-HEPES+10% FCS, TCM-HEPES+rPF(v/v, 50/50), or rPE only and examined the nuclear maturation after aceto-orcein or hochest staining. After identifying the optimal incubation time, this experiment was repeated for 5 times. Under the TCM-HEPES containing hormones and serum codition, the time range of porcine follicular oocyte maturation was 38 to 44 hours and the optimal time of maturation of follicular oocyte in vitro was 42 hour cultivation, respectively. The maturatin rates(89.4% and 92.7%) of porcine follicular oocytes cultured in the media with 50% rPF or only rPF were signifciantly higher thanthat (84.6%) of oocytes cultured with TCM-HEPES, respectively. These results suggest that the unknown components(s) of rPF promoted in vitro maturation of porcine follicular oocytes.

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Reproductive fecundity of Iraqi Awassi ewes immunized against synthetic inhibin-α subunit or steroid-free bovine follicular fluid

  • Al-Sa'aidi, Jabbar Abbas Ahmed;Khudair, Khalisa Khadim;Khafaji, Sura Safi
    • Asian-Australasian Journal of Animal Sciences
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    • v.31 no.8
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    • pp.1169-1175
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    • 2018
  • Objective: The present study was conducted to investigate the impacts of active and passive immunization against synthetic inhibin and steroid-free bovine follicular fluid, respectively, on reproductive fecundity out of breeding season in Iraqi Awassi ewes. Methods: Follicular fluid was aspired from mature bovine follicles, treated with activated charcoal, and used for immunization of male rabbits for obtaining steroid free bovine follicular fluid (SFBFF) antiserum. Forty non-pregnant Awassi ewes were allocated into 4 groups (n = 10 each). At day 38 of experiment, ewes were treated with intra-vaginal MPA sponge (60 mg for 12 days). At days 0, 28, and 50, ewes were treated with 4, 2, and 2 mL of normal saline (control; C-ve), 400, 200, and $200{\mu}g$ of ovalbumin (C+ve), 400, 200 and $200{\mu}g$ of inhibin (SI group), respectively, and 4 mL of normal saline at day 0, and 4 and 2 mL of SFBFF antiserum at days 28 and 50, respectively, (AI group). After mating with Awassi rams, pregnancy and embryo number were diagnosed, at day 38 of pregnancy, using ultrasonography. Blood samples were collected at days 30, 60, 90, and 120 of pregnancy, for assessment of estradiol-$17{\beta}$ (E2) and progesterone (P4) levels. After parturition, numbers of delivered lambs were recorded. Results: The results revealed significant increase of P4 and significant decrease of E2 levels in SI and AI pregnant ewes than controls at days 30, 60, 90, and 120. Newborn number increased significantly in SI and AI treated than control ewes. Conclusion: Active or passive immunization against endogenous inhibin could augment reproductive fecundity out of breeding season in Iraqi Awassi ewes.

Influence of Human Follicular Fluid and Gonadotropins in the Culture Medium on the In Vitro Maturation, Fertilization and Development of Human Immature Oocytes (배양액 내 인간 난포액 및 성선자극호르몬 첨가가 인간 미성숙 난자의 체외성숙, 수정 및 체외 배발달에 미치는 영향)

  • Kim, Eun-Kuk;Kim, Dong-Won;Jeong, Byung-Jun
    • Journal of Embryo Transfer
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    • v.24 no.3
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    • pp.145-150
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    • 2009
  • This study was conducted to examine the effects of human follicular fluid and gonadotropin (FSH+HCG+rhEGF) on in vitro maturation, fertilization and development of human immature oocytes. Cumulus-oocyte complexes (COCs) were collected following for in vitro fertilization and embryo transfer (IVF-ET) cycles of the patients. At the time of oocytes collection, oocytes were classified into MII, MI and GV in accordance with their appearance (MII: Fully mature oocyte at metaphase II of meiosis; MI: Nearly mature oocytes at metaphase I of meiosis; GV: Immature oocytes at prophase I of meiosis). After controlled ovarian stimulation using gonadotropin(FSH) and human chorionic gonadotropin (HCG) in 70 ICSI cycles, 158 MI to MII matured oocytes were intracytoplasmic sperm injection (ICSI) ${\sim}4$ h after in vitro culture and 553 MII oocytes were ICSI after denudation. The aspirated MI and GV oocytes were cultured in culture medium containing 10% (v/v) serum protein substitute (SPS), 10% (v/v) human follicular fluid (hFF) and 10% (v/v) serum protein substitute (SPS)+1 IU/ml FSH+10 IU/ml HCG+10 ng/ml recombinant human epidermal growth factor (rhEGF). The maturation rate of immature oocytes was similar among the three group. When maturation medium was supplemented with 10% SPS, 10% hFF or gonadotropins, the fertilization rate of in vitro matured oocytes was higher in 10% SPS (80.0%), but there was no statistical significance (78.2%; hFF, 76.9%; gonadotropin, p>0.05). The development rate of human embryos developed to $6{\sim}8$ cells were not significant difference in the medium containing SPS, hFF and gonadotropins (65.6%, 65.9% and 66.7%). The results of these study suggest that human follicular fluid and gonadotropins supplemented in the culture medium was not effected on the in vitro maturation, fertilization and development of human immature oocytes.