• Title/Summary/Keyword: Reproductive Fluid

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Studies on the Suitability and Efficiency of Human Follicular Fluid as Protein Supplement in Assisted Reproductive Technology(ART);II. Effect of Human Follicular Fluid on Development of Mouse Embryos In Vitro (생식보조시술시 단백질원으로서 인간난포액의 적합성 및 효율성에 관한 연구;II. 인간난포액이 생쥐수정란의 체외발달에 미치는 효과)

  • Kim, D.H.;Chi, H.J.;Kim, J.Y.;Koo, J.J.;Chang, S.S.;Chung, K.S.
    • Clinical and Experimental Reproductive Medicine
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    • v.23 no.1
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    • pp.95-102
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    • 1996
  • This study was performed to investigate the effect of human follicular fluid (HFF) on development of mouse embryos, for evaluating the suitability of HFF as a substitutive material of human fetal cord serum in ART program. The various concentrations of HFF were added into the culture medium and the effects of HFF concentrations were examined to identify the optimal concentration of HFF for embryo development. The potency of HFF in improving embryo development was compared to that of other protein supplement. Collected HFFs were classified with the maturity of the containing oocytes; mature, immature, atretic, and then the effects of the classified HFFs on embryo development were examined. Also, HFF was separated into the low (<30,000 Da) and high (>30,000 Da) molecular weight fractions and the effects of the fractions on embryo development were investigated. The highest development rate was found in culture medium supplemented with 20% HFF, bnt this rate was reversely reduced at the concentrations of HFF higher than 20%. The development rates to the blastocyst, hatching blastocyst, attachment and outgrowth cultured in mature HFF was significantly higher than those in immature and atretic HFF, and mean cell number in blastocyst was higher in mature HFF than in immature and atretic HFF. The development rates of mouse embryos according to protein sources were significantly higher in HFF than in fetal cord serum (FCS), maternal serum (MS) and bovine serum albumin (BSA), and mean cell number in blastocyst cultured in HFF was higher than that in FCS, MS and BSA. The development rates of embryo and mean cell number in blastocyst cultured in high molecular weight fraction of HFF were higher than those in low molecular weight fraction, but the results of high molecular weight fraction were lower than those of whole HFF. Therefore, these results indicated that human mature follicular fluid was useful for improving the development of mouse embryos, which suggests a possibility that HFF also may be used efficiently for improving the culture condition in human ART program as a protein supplement.

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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.

Understanding of Intrauterine Environment Changes based on Proteomics and Bioinformatics during Estrous Cycle (단백체학과 생물정보학을 이용한 자궁 내 환경의 이해)

  • Lee, Sang-Hee;Lee, Seunghyung
    • Journal of Life Science
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    • v.29 no.5
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    • pp.621-630
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    • 2019
  • Fertilization is the beginning of a new life that occurs in the female uterine. The female reproductive tract is composed ovary, oviduct, uterine, vagina and cervix, their physiological features are regulated by estrous cycle. Of these, uterine is a main point to establish embryo development and implantation, and intercommunication between embryo and uterine environment is necessary for suitable pregnancy. Endometrium is part of the uterine, its morphology is repetitively changed by hormones, and characteristic of uterine fluid from endometrium is also changed. Recently, massive proteins of endometrium and uterine fluid can be detected according to develop proteomics and bioinformatics and have been accelerated the understanding of the reproductive biology fields. Moreover, the massive protein information is actively studying with deeply studied theory such as sex hormone signal pathway and angiogenesis in mammals. In this paper, we review understanding of endometrium remodeling, uterine gland and fluid during estrous cycle, additionally studies on endometrium and uterine fluid based on proteomics techniques. Lastly, we introduced methods of the protein-protein correlation using bioinformatics tool that interaction with hormone receptors, representative angiogenetic factors and detected proteins using proteomics in endometrium and uterine fluid. This review will be useful to understanding the study on search of new cell mechanism in endometrium and uterine fluid.

Comparison of PRRSV and antibody detection in oral fluid and serum samples from different age categories of PRRSV endemic farms (PRRS 양성농장의 사육단계별 구강액과 혈액을 이용한 PRRSV와 항체 검출 비교)

  • Kim, Jung-Hee;Son, Jae Guk;Kim, Won-Il
    • Korean Journal of Veterinary Service
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    • v.43 no.3
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    • pp.173-179
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    • 2020
  • The objective of this study was to evaluate the usefulness of detection of PRRSV and PRRSV-specific antibodies in oral fluids for monitoring of PRRSV infection in endemic farms. The level of PRRSV and anti-PRRSV antibodies in serum and oral fluids was evaluated in five age groups of pigs (6, 9, 12, 16 weeks of age and gilts). The samples (25 serums and 5 oral fluids/per a farm) were collected from 5 different farms endemically infected by PRRSV. Both serum and oral fluid samples were tested for PRRSV by quantitative reverse-transcription polymerase chain reaction (qRT-PCR) and for anti-PRRSV antibodies by two commercial PRRSV ELISA kits. ELISA mean s/p ratios (2.98 vs 1.63) and positive rate (84.0% vs 68.8%) of the oral fluid samples showed significantly higher levels but had similar patterns to the seroprofile of the blood samples. The PRRSV positive rate of oral fluid and serum samples was 40.0% and 44.0% respectively. In conclusion, the use of oral fluids for PRRS monitoring in endemic farms is strongly recommended.

Effect of Hydrosalpingeal Fluid on the Implantation in-vitro in a Murine Model (생쥐 배아의 체외배양 시 착상과정에 대한 난관수종액의 영향)

  • Jun, Jin-Hyun;Koong, Mi-Kyoung;Lim, Chun-Kyu;Kim, Soo-Kyung;Kang, Inn-Soo
    • Clinical and Experimental Reproductive Medicine
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    • v.27 no.2
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    • pp.159-164
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    • 2000
  • 연구목적:인간의 체외수정 및 배아이식술에서 난관수종을 갖는 환자에서 임신율과 착상률이 감소된다는 보고들이 있지만 이에 대한 명확한 기작은 밝혀지지 않았다. 본 연구에서는 생쥐 배아를 이용한 체외 착상모델에서 인간의 난관수종액(HSF)이 착상과정에 미치는 영향을 알아보고자 하였다. 연구재료 및 방법 난관수종액은 난관수종으로 수술을 받은 8명의 환자로부터 채취하였으며, 실험에 사용하기 전까지 냉동고에 보관하였다. 생쥐의 포배기 배아는 2-세포기배아를 3일 동안 배양하여 그 중 상태가 양호한 포배기 배아만을 선별하여 투명대를 제거한 후 사용하였다. 기본 배양액으로는 Ham's F-10을 사용하였으며, 배양 시 기본 배양액만을 사용한 경우를 group Ⅰ으로 하였고, 기본 배양액에 0.5% FBS를 첨가한 경우를 group Ⅱ, 0.5% FBS와 50% HSF를 첨가한 경우를 group Ⅲ , 100% HSF에 0.5% FBS를 첨가한 경우를 group Ⅳ,100% HSF만을 사용한 경우를 group Ⅴ로 하였다. 투명대를 제거한 포배기 배아를 각각의 HSF에 대한 5종류의 배양액에서 48시간 동안 배양하였다. 체외 착상 유무는 부착 부위에서 크기가 커진 영양세포들을 관찰하여 판정하였으며, 착상 부위의 표면적은 화상분석기를 이용하여 산출하였다. 결 과: 생쥐 배아의 체외 착상률은 group Ⅰ, Ⅱ, Ⅲ, Ⅳ, Ⅴ에서 각각 0%, 98.9%, 77.5%, 40.4%, 10.0%로 나타났으며, 착상 부위의 평균 표면적은 group Ⅱ, Ⅲ, Ⅳ, Ⅴ에서 각각 $74,675{\pm}25,201{\mu}m^2$, $59,024{\pm}25,877{\mu}m^2$, $45,156{\pm}22,654{\mu}m^2$, $38,254{\pm}17,115{\mu}m^2$이었다. 체외 착상률과 부위의 표면적은 HSF의 농도가 증가함에 따라 통계적으로 유의하게 감소하였다(p<0.001). 결론:인간의 난관수종액(HSF)은 생쥐 배아의 체외 착상과 영양배엽세포의 증식을 억제하는 것으로 확인되었으며, 이러한 원인이 난관수종을 갖는 환자에서 임신율이 낮은 것과 밀접한 관련이 있을 것으로 생각된다.

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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.

Effect of Mature Human Follicular Fluid on the Development of Mouse Embryos in vitro (성숙난포액을 이용한 생쥐배아의 발달에 관한 연구)

  • Park, S.Y.;Lee, J.J.;Kim, S.H.;Ku, P.S.
    • Clinical and Experimental Reproductive Medicine
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    • v.19 no.2
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    • pp.125-131
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    • 1992
  • The possible effect of human follicular fluid(hFF) on the growth and development of fertilized oocytes and embryos is important because the fallopian tubes are exposed to FF after follicular rupture and the processes of fertilization and embryo cleavage occur inside the fallopian tubes. Previously, it was suggested that human FF might adversely affect on the development of early mouse embryos. In order to investigate the effect of hFF on the development of embryos, early mouse embryos were cultured in media containing various protein sources as bovine serum albumin(BSA), fetal cord serum(FCS) and FF. And we evaluated the development of early mouse embryos in terms of the morphology, cleavage rate, and cell count of blastcysts. There were no significant differences in the morula and blstocyst formation rates of 2-cell mouse embryos cultured in the media containg three different protein sources and three different concentrations of FF. The blastocyst formation rate of 1-cell mouse embryo cultured in FF group was significantly higher than that cultured in BSA group(P<0.05). The morula and blastocyst formation rates of 2-cell mouse embryos of the group cultured in the media containing FF were comparable with those of other two groups, in addition, the cell count of blastocysts of FF group in the 2-cell embryo culture was higher than those of BSA group and HCS group(P<0.01), and this finding was also noted in 1-cell embryo culture. There was no difference in the morula and blastocyst formation rates of the 2-cell mouse embryos cultured in the media containing different concentrations of FF. These results suggest that mature human follicular fluid has no inhibitory activity on the development of early mouse embryos even in high concentration and may be a good protein source which is positively associated with the development of mouse embryos in vitro especially in 1 cell embryo culture.

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The Effect of Antisperm Antibodies Detected by Immunobead Binding Assay on Fertilization and Cleavage of Human Oocytes In Vitro (Immunobead 검사로 검출된 항정자 항체가 인간 난자의 체외 수정 및 분할에 미치는 영향)

  • Chung, Dong-Geun;Shin, Chang-Jae;Moon, Shin-Yong;Chang, Yoon-Seok
    • Clinical and Experimental Reproductive Medicine
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    • v.16 no.2
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    • pp.153-160
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    • 1989
  • The effect of antisperm antibodies (ASA) on the human in vitro fertilization (lVF) process was evaluated by analyzing the IVF data between October and December 1988 at Seoul National University Hospital prospectively. The immunobead test (IBT) was used to identify Ig G, Ig A, and Ig M in the serum, semen, and follicular fluid from 93 couples undergoing in vitro fertilization-embryo transfer (lVF-ET ) . The fertilization rate in couples with ASA to sperm head of at least one isotype in female serum (n= 10) was significantly less than that in couples without ASA to sperm head (n=83; 28.5% versus 45.3% , p=0.028). The presence of ASA to sperm head in follicular fluid (n=8) also reduced fertilization rate from 45.3% to 24.4% (p=O.0l3). However, ASA binding to sperm head in male serum and semen did not predict fertilization. Similarly, ASA binding to sperm tail and tail-tip did not reduced the oocyte fertilization rate significantly in any of the fluids tested. The zygote cleavage rate was not reduced in the presence of ASA. These results suggest that the presence of ASA to sperm head in female serum and follicular fluid is associated with reduced fertilization in IVF-ET. Another observation is that the oocyte that do fertilize in the presence of antisperm antibodies can subsequently proceed with normal cleavage. The results of this investigation therefore suggest that the IBT is a useful test forscreening of women participat.ing IVF-ET program.

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Lower growth factor expression in follicular fluid undergone in-vitro fertilization

  • Han, Myoung-Seok;Park, Seung-Bin;Park, Bang-Ja
    • Clinical and Experimental Reproductive Medicine
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    • v.38 no.4
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    • pp.210-215
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    • 2011
  • Objective: This study was performed to identify whether growth and differentiation factor-9 (GDF-9) and transforming growth factor-${\beta}1$ (TGF-${\beta}1$) expressions would be lower in the follicular fluid (FF) of those over age 35 who underwent IVF than under age 35. Methods: A total of 24 IVF cycles (20 patients) were included in this study. All of patients were stimulated for IVF by the GnRH short protocol and divided into two groups for analysis, according to their age: <35 group (14 cycles, 11 patients) vs. ${\geq}35$ group (10 cycles, 9 patients). The expression levels of GDF-9 and TGF-${\beta}1$ were determined by western blotting and quantitative enzyme-linked immunosorbent assay. Results: The numbers of retrieved oocytes and metaphase II oocytes were significantly lower in the ${\geq}35$ group. Lower expression of GDF-9 and TGF-${\beta}1$ by western blotting in the ${\geq}35$ group were observed as well. The mean GDF-9 and TGF-${\beta}1$ levels by enzyme-linked immunosorbent assay were lower in the ${\geq}35$ group. The values were $6,850.5{\pm}928.4$ ng/L vs. $3,333.3{\pm}1,089.2$ ng/L of GDF-9 ($p$ <0.05) and $3,844.1{\pm}571.1$ ng/L vs. $2,187.7{\pm}754.0$ ng/L of TGF-${\beta}1$ ($p$ <0.05). A negative correlation between GDF-9 and age was observed (r=-0.546, $p$=0.006). Conclusion: GDF-9 and TGF-${\beta}1$ production from stimulated ovaries during IVF appears to decrease with age.

Relationship between follicular fluid adipocytokines and the quality of the oocyte and corresponding embryo development from a single dominant follicle in in vitro fertilization/intracytoplasmic sperm injection cycles

  • Chang, Hye Jin;Lee, Ji Hyun;Lee, Jung Ryeol;Jee, Byung Chul;Suh, Chang Suk;Kim, Seok Hyun
    • Clinical and Experimental Reproductive Medicine
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    • v.41 no.1
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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.