294 THE EFFECT OF PLASMIN ON THE IN VITRO FERTILIZATION ABILITY OF PORCINE GAMETES

2006 ◽  
Vol 18 (2) ◽  
pp. 254
Author(s):  
H.-H. Rhee ◽  
S.-J. Sa ◽  
H.-T. Cheong ◽  
B.-K. Yang ◽  
C.-K. Park
Keyword(s):  
In Vitro Fertilization ◽  
Zona Pellucida ◽  
Acrosome Reaction ◽  
Proteolytic Enzymes ◽  
Extracellular Proteins ◽  
Control Group ◽  
Sperm Viability ◽  
Vitro Fertilization ◽  
Sperm Binding

Plasminogen activators (PAs) are specific proteolytic enzymes that convert the inactive proenzyme plasminogen to plasmin. The plasmin formed is a nonspecific, potent protease that cleaves blood fibrin clots and several other extracellular proteins. The purposes of the present study were (1) to assess the effect of plamin on sperm viability and acrosome reaction (AR), (2) to examine the effect of plasmin on zona pellucida (ZP) solubility and the binding of sperm to ZP, and (3) to evaluate the effect of plasmin on fertilization responses, including penetration and incidence of polyspermy during in vitro fertilization in the pig. Ejaculated semen was collected from three mature Duroc boars by artificial vagina. The same three boars were used for all experiments. The oocyte maturation medium used was North Carolina State University-23 (NCSU-23) medium supplemented with 10% (v/v) porcine follicular fluid (pFF), 0.6 mM cysteine, 10 IU/mL human chorionic gonadotropin (hCG; Sigma-Aldrich Corporation, St. Louis, MO, USA), and 10 IU/mL pregnant mare's serum gonadotropin (PMSG; Sigma). Porcine spermatozoa, which were washed in Dulbecco PBS (Sigma), were resuspended and incubated in fertilization medium (mTBM) containing 0, 0.1, 1.0, 10.0, or 100.0 ng/mL plasmin (Sigma). Data were analyzed by ANOVA and Duncan's multiple-range test using the Statistical Analysis System (SAS Institute, Inc., Cary, NC, USA). The present study suggests that sperm viability was not affected by plasmin treatment. Also, addition of plasmin in doses ranging between 0.1 and 100.0 ng/mL for 2, 4, or 6 h to washed boar spermatozoa resulted in enhancement of acrosome reaction (AR), compared with untreated cells. Concentrations of 0 and 0.1 ng/mL plasmin (83 � 15 and 95 � 18 sperm/oocyte, respectively) had no effect on sperm binding, whereas 1.0 (123 � 21 sperm/oocyte), 10.0 (124 � 16 sperm/oocyte), and 100 ng/mL (124 � 15 sperm/oocyte) plasmin increased (P < 0.05) sperm binding, compared with the control. The zona pellucida solubility (zona digestion time) was significantly (P < 0.05) lower in medium with 1.0 (123 � 24 s), 10.0 (99 � 15 s), or 100.0 ng/mL (95 � 19 s) plasmin, compared with control (176 � 27 s). When porcine oocytes and spermatozoa were co-incubated in various concentrations of plasmin for 6 h, the penetration rate was significantly (P < 0.05) higher in medium with 1.0 ng/mL plasmin (77.5 � 3.1%), compared with control. However, there were no significant differences in the polyspermic rates and mean numbers of sperm (MNS)/oocyte among the groups treated with plasmin and the control group. We found that addition of plasmin to fertilization medium increases the percentage of acrosome-reacted spermatozoa and the sperm-binding ability of the pig ZP. These results suggest that plasmin may play a role in events related to fertilization in the pig.

Differences between antigenic determinants of pig and cat zona pellucida proteins

Reproduction ◽  
2000 ◽  
pp. 15-23 ◽  
Author(s):  
K Jewgenow ◽  
M Rohleder ◽  
I Wegner
Keyword(s):  
In Vitro Fertilization ◽  
Zona Pellucida ◽  
Antigenic Determinants ◽  
Vitro Fertilization ◽  
Contraceptive Vaccine ◽  
Sperm Binding ◽  
Dependent Inhibition ◽  
Dose Dependent Inhibition ◽  
Dose Dependent

Despite many efforts, the control of reproduction in feral cat populations is still a problem in urban regions around the world. Immunocontraception is a promising approach; thus the present study examined the suitability of the widely used pig zona pellucida proteins (pZP) for contraception in feral domestic cats. Purified zona pellucida proteins obtained from pig and cat ovaries were used to produce highly specific antisera in rabbits. Antibodies against pZP raised in rabbits or lions were not effective inhibitors of either in vitro sperm binding (cat spermatozoa to cat oocytes) or in vitro fertilization in cats, whereas antibodies against feline zona pellucida proteins (fZP) raised in rabbits showed a dose-dependent inhibition of in vitro fertilization. Immunoelectrophoresis, ELISA and immunohistology of ovaries confirmed these results, showing crossreactivity of anti-fZP sera to fZP and to a lesser extent to pZP, but no interaction of anti-pZP sera with fZP. It is concluded that cat and pig zonae pellucidae express a very small number of shared antigenic determinants, making the use of pZP vaccine in cats questionable. A contraceptive vaccine based on feline zona pellucida determinants will be a better choice for the control of reproduction in feral cats if immunogenity can be achieved.

Relationship between zona pellucida–induced acrosome reaction, sperm morphology, sperm–zona pellucida binding, and in vitro fertilization

2003 ◽  
Vol 79 (1) ◽  
pp. 49-55 ◽  
Author(s):  
Hadley S Bastiaan ◽  
Mari-Lena Windt ◽  
Roelof Menkveld ◽  
Thinus F Kruger ◽  
Sergio Oehninger ◽  
...  
Keyword(s):  
In Vitro Fertilization ◽  
Zona Pellucida ◽  
Acrosome Reaction ◽  
Sperm Morphology ◽  
Vitro Fertilization

Levels interleukine-4 and interleukin-17 (IL-4, IL-17) of blood in patients with habitual recurrent pregnancy loss, which occurred in a loop in vitro fertilization

2016 ◽  
pp. 137-139
Author(s):  
K.P. Golovatyuk ◽  
Keyword(s):  
In Vitro Fertilization ◽  
Conditioned Medium ◽  
Mononuclear Cells ◽  
Recurrent Miscarriage ◽  
Interleukin 4 ◽  
Control Group ◽  
Interleukin 17 ◽  
Vitro Fertilization ◽  
Blood Mononuclear Cells

The objective: was to investigate the levels of cytokines IL-4 and IL-17 in serum and conditioned medium cultures of blood mononuclear cells (MNC) and evaluation association between their products and miscarriage, which occurred in IVF cycles. Patients and methods. We observed 240 patients with recurrent miscarriage, came in IVF cycles, and 100 apparently healthy fertile women in the control group. The concentrations of IL-4 and IL-17 in serum and conditioned medium of MNC cultures were determined. Results. The levels of IL-4 in the serum and conditioned medium in spontaneous and stimulated mitogen secretion was not significantly different from those in the control group, whereas IL-17 levels were higher than those in the control group serum, in conditioned media of stimulated and non-stimulated MNCs. Conclusion. Disregulation of activity of circulating blood mononuclear cells in women with recurrent miscarriage that followed IVF, is accompanied by increased secretion of IL-17 and almost constant production of IL-4 on the back of high stimulation index of production of these cytokines. Key words: in vitro fertilization, miscarriage, interleukin-4, interleukin-17, serum stimulated and non-stimulated mononuclear blood.

scholarly journals IN VITRO FERTILITY TEST OF HUMAN SPERMATOZOA MEMBRANE PROTEIN FERTILIN BETA ANTIBODY IN MICE (Mus musculus Balb/c) AS IMMUNOCONTRACEPTIVE CANDIDATE

2017 ◽  
Vol 52 (3) ◽  
pp. 209
Author(s):  
Reny I’tishom ◽  
Doddy M Soebadi ◽  
Aucky Hinting ◽  
Hamdani Lunardhi ◽  
Rina Yudiwati
Keyword(s):  
In Vitro Fertilization ◽  
Membrane Protein ◽  
Mus Musculus ◽  
Human Spermatozoa ◽  
Control Group ◽  
Reproductive Process ◽  
Vitro Fertilization ◽  
Antibody Induction

One of the materials as potential candidates immunocontraception material is spermatozoa. Fertilin beta is spermatozoa membrane protein and is found only in mature spermatozoa and ejaculate, which serves as an adhesion molecule. Spermatozoa membrane protein that is used as an ingredient immunocontraception candidate, must have specific criteria that the specificity of spermatozoa, the role of antigen in the fertilization process, which includes the formation of immunogenicity sufficient antibody response has the potential to block fertilization. Antibodies against spermatozoa affect the stages before fertilization of the reproductive process and can hinder the development of the embryo after fertilization. Until now very little research data spermatozoa membrane protein as an ingredient immunocontraception are up to the test of experimental animals. The research objective is to prove the role of the resulting antibody induction of antibodies fertilin beta protein in the membrane of human spermatozoa induce agglutination and reduce motility thus reducing the number of in vitro fertilization. Research conducted at the IVF Laboratory, Department of Biology of Medicine, Faculty of Medicine, University of Airlangga. This research includes: Test the potential of antibody protein beta fertilin membrane of human spermatozoa and inhibit the role of antibodies in vitro fertilization in mice (Mus musculus Balb/c). In vitro studies have resulted in fertilization figure of 25% is smaller than the number that is equal to control fertilization of 58.7%, whereas previously the spermatozoa were incubated first with a beta membrane protein antibody fertilin human spermatozoa. While the percentage of inhibition of sperm to fertilize an oocyte by 33.75%. Potential imunokontraseptif considered effective if it decreased significantly (P <0.05) than the numbers fertilization in the treatment group compared with the control group. This shows fertilin beta membrane protein antibody has the ability to inhibit human spermatozoa to fertilize oocytes that reduce the number of fertilization.

scholarly journals The effectiveness of collaborative infertility counseling (CIC) on pregnancy outcome in infertile women undergoing in vitro fertilization: A randomized controlled trial

2019 ◽  
Author(s):  
Mahboobeh Rasoulzadeh Bidgoli ◽  
robab latifnejad roudsari ◽  
ali montazeri
Keyword(s):  
In Vitro Fertilization ◽  
Pregnancy Rate ◽  
Treatment Success ◽  
Intervention Group ◽  
Control Group ◽  
Vitro Fertilization ◽  
Infertile Women ◽  
Significant Difference ◽  
And Control

Abstract Background: Infertility is an emotional tension which influences the whole aspects of relationships in infertile couples. A main objective of infertility treatments is elevation of pregnancy rate. The present study aimed to examine the effect of collaborative counseling on pregnancy rate in infertile women, undergoing in vitro fertilization in Mashhad, Iran. Methods: In this clinical trial, 60 women with primary infertility were selected from an infertility research center and were randomly allocated into intervention (n=29) and control (n=31) groups. The intervention group received individual counseling, based on the collaborative reproductive healthcare model with collaboration of a midwife, a gynecologist and a clinical psychologist in five sessions during a two-month period. The control group received routine care. Positive pregnancy test was considered as a criterion of treatment success at the end of the study. Data were analyzed using statistical tests including independent samples t-test. Results: There was no significant difference in pregnancy rate between intervention and control groups (P = 0.298). Also, there were no significant differences in follicle and embryo numbers between two groups. However, a significant difference was observed between two groups in terms of oocyte numbers where the intervention group had more oocyte (P = 0.014). Conclusion: Overall the findings indicated that the collaborative infertility counseling did not improve treatment success in infertile women undergoing in vitro fertilization

scholarly journals Piglets produced by transfer of embryos obtained by in vitro fertilization of oocytes matured in vitro with thymosin: A case report

2020 ◽  
Vol 76 (03) ◽  
pp. 6356-2020 ◽  
Author(s):  
KATARZYNA PONIEDZIAŁEK-KEMPNY ◽  
BARBARA GAJDA ◽  
IWONA RAJSKA ◽  
LECHOSŁAW GAJDA ◽  
ZDZISŁAW SMORĄG
Keyword(s):  
Case Report ◽  
In Vitro Fertilization ◽  
Control Group ◽  
First Birth ◽  
Vitro Fertilization ◽  
Research Material ◽  
Preliminary Study ◽  
Experimental Group

The aim of the study was to examine the in vivo viability of in vitro-produced (IVP) porcine embryos obtained from oocytes matured with thymosin. The research material for this study consisted of immature pig oocytes obtained from ovaries after slaughter and ejaculated semen obtained from one boar. The immature oocytes were cultured in vitro until the metaphase II stage in a medium supplemented with thymosin (TMS). The presumptive zygotes obtained were cultured in vitro for 4-40 hours. The presumptive zygotes and 2-4-cell embryos were evaluated in vivo after transferring them to synchronized recipients. After the transfer of embryos from the experimental group into 2 recipients (50 embryos into each gilt) and the transfer of 50 embryos from the control group into 1 recipient, both gilts that had received embryos obtained by in vitro fertilization of oocytes matured with TMS became pregnant and delivered a total of 16 live piglets. After the transfer of embryos from the control group, no pregnancy was achieved. In conclusion, the results of our preliminary study suggest that the maturation of pig oocytes with thymosin supports the in vivo survival of in vitro produced embryos. It is important to note, that this was the first birth of piglets obtained after transfer of IVP embryos in Poland.

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