• Korean Journal of Animal Reproduction
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• v.20 no.2
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• pp.127-134
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• 1996

The objective of this experiment was to test the ability of the fertilization of EGF treated pig oocytes for in vitro maturation. The addition of EGF (10 ng/ml), FSH (10 ${\mu}\textrm{g}$/ml), or FBS (10%) on maturation medium of pig immature oocytes divided into four groups as follows; group 1: untreatment, group 2: EGF alone, group 3: combination of FSH and FBS, or group 4: combination of EGF, FSH, and FBS. The interactive effects of nuclear maturation rates (M II%) of EGF alone, FSH plus FBS, and EGF plus FSH added FBS treatments were significantly higher than those of non-treatments (P<0.001). The fertilization rate of EGF alone (group 2) was lower than that of 3, 4 groups, but was significantly higher than group 1 (p< 0.005). Furthermore, combination of EGF, FSH,and FBS (group 4) was higher than others (group 1. 2, 3) on male pronuclei formation as well as penetration of sperm (P<0.05). These results suggested that EGF alone decreased the ability of cytoplasmic maturation compared to nuclear maturation in pig oocytes, but a high level of cytoplasmic maturation of in vitro-matured pig oocytes can be achieved when supplemented with FSH and FBS.

• Korean Journal of Animal Reproduction
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• v.24 no.2
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• pp.143-154
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• 2000

Most eggs initiated the fertilization processes but arrested at specific stages. The stages included failure of the oocyte to exit from the meiotic metaphase-II with or without sperm penetration, failure of appropriate sperm aster formation, inability to form proper male and female pronuclei, failure of suitable pronuclear apposition, and failure to form proper number of either male or female pronuclei. Various images of defective microtubule organization and chromatin configuration during IVF and ICSI procedures were observed. We discussed the data with previous research results during normal fertilization in humans and other mammals. In conclusion, various aberrant patterns in microtubule assembly and chromatin configuration, which were assessed in the present study, could be used as criteria to improve assisted reproductive technology in clinics. However, further cellular and molecular characterization is needed to clarify these aberrant patterns of cytoskeletal assembly.

• Korean Journal of Animal Reproduction
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• v.17 no.3
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• pp.233-242
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• 1993

The effects of in vitro maturation and sperm treatment condition on the in vitro fertilization (IVF) and developmental capacity of bovine oocytes were investigated and the development of embryos was compared under the 2 different co-culture system, with GC or BOEC. The cultured embryo to 16 cell or morula wre transferred into recipients or frozen by 2 different freezing method. The results obtained were summarized as follows; 1. In vitro maturation rates of vovine follicular oocytes cultrued in TCM199 with 10% FCS or ECS were 64.0% and 72.7%, but the case of addition of 10% FCS or ECS to TCM199 co-cultured with granulosa cells were 81.3% and 84.0%, respectively. IVM rate of three TCM199 added to granulosa cells was higher than that of media without granulosa cells. 2. When bovine follicular oocytes were matured in TCM199 with 10% FCS and GC and then fertilized in vitro by sperm treated with caffeine, embryo developments of bovine oocytes co-cultured with BOEC were 38.4% and 51.4%, respectively. But those of bovine oocytes co-cultured with GC were 52.2% by sperm treated with caffeine-heparin. 3. Cleavage rates of bovine oocytes cultured with 10% FCS alone and fertilized in vitro by sperm treated with caffeine-heparin was 33.0%. 4. When bovine follicular oocytes were matured in TCM199 with 10% FCS and GC, embryo developments of bovine ooctyes co-cultured with BOEC of GC were 46.0% and 50.2%, respectively. 5. When bovine follicular oocytes were matured in TCM199 with 10% ECS and GC, embryo developments co-cultured with BOEC or GC were 45.2% and 51.4%, respectively. 6. When Korean Native cow's follicular oocytes matured in TCM199 with 10% FCS and GC, embryo developed co-cultured with BOEC or GC were 45.2% and 51.4%, respectively. 6. When Korean Native cow's follicular oocytes matured in TCM199 with 10% FCS and GC, embryo developments of the bovine oocyte co-cultured with BOEC and GC were 41.8% and 60.1%. But with FCS 10% those of the bovine oocytes co-cultured with BOEC and GC were 42.0% and 48.4%, respectively. 7. When Holstein's follicular oocytes were matured in TCM199 with 10% ECS and GC, embryo developments fo the bovine oocytes co-cultured with BOEC and GC were 50.0% and 57.7%, but with ECS 10% those of the bovine oocytes co-cultured with BOEC and GC were 52.2% and 56.5%, respectively. 8. The viability of frozen-thawed embryos ranged from 60~80% and those of frozen-thawed embryos from vitrification was lower than that from conventional metiod. 9. The selected fresh embryos were transferred nonsurgically to 7 recipients but did not result in pregnancy.

• Clinical and Experimental Reproductive Medicine
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• v.48 no.2
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• pp.142-149
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• 2021

Objective: Bacteriospermia and urogenital infections are common problems in male infertility. This study aimed to evaluate the effects of bacteriospermia on sperm parameters and clinical outcomes in semen samples infected with two common bacteria (Staphylococcus saprophyticus and Escherichia coli) in northern Iran. Methods: Microbiological tests were performed to isolate and identify organisms from 435 semen samples from infertile couples. Semen samples were assessed according to the World Health Organization criteria. The protamine status, chromatin structure, chromatin condensation, and acrosome reaction of sperm and assisted reproductive outcomes were determined in couples with different male infertility factors. Results: Among the total cases, the two most prevalent pathogens were considered: S. saprophyticus (38.2%) and E. coli (52.9%). In the semen samples infected with E. coli, the spontaneous acrosome reaction and abnormal chromatin condensation were more common (p<0.05). Significant increases in abnormal chromatin condensation and deprotamination were seen in the presence of S. saprophyticus. In washed semen, tight adhesion between the sperm midpiece and S. saprophyticus was observed. There was also a significant decrease in the fertilization rate using semen samples infected with S. saprophyticus and E. coli during in vitro fertilization cycles (p<0.001). In addition, the presence of S. saprophyticus and E. coli in semen samples was associated with a lower likelihood of clinical pregnancy in couples with various factors of male infertility. Conclusion: Poor results of assisted reproductive techniques may be correlated with semen samples infected with two common bacteria in northern Iran.

• Reproductive and Developmental Biology
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• v.38 no.1
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• pp.53-62
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• 2014

The objective of this study was to investigate the relationship between fertility and protein pattern change using in vitro fertilization, analysis of sperm characteristics and two-dimensional gel electrophoresis in different pig types. In results, the viability and mitochondria integrity of sperm were higher significantly (p<0.05) but the portions of acrosome reaction was lower significantly (p<0.05) in Duroc and $F_1$ (potbellied ${\times}$ PWG miniature pig) than PWG miniature. On in vitro fertilization to investigate fertility, the fertility of $F_1$ semen war higher significantly (p<0.05) than in Duroc and PWG miniature pig. On the other hand, protein patterns showed similar function among the different boar semen. Especially, the heat shock 70 kDa 1-like and G patch domain-containing protein 4 were significantly (p<0.05) higher expressed in $F_1$ than in Duroc and PWG miniature pig. The proteins associated with mitochondria in Duroc were significantly (p<0.05) higher expressed than in $F_1$ and PWG miniature pig. The developmental rates to blastocyst stage of oocytes fertilized with sperm of $F_1$ pig were significantly (p<0.05) higher than in PWG miniature pig. However, phosphoglycerate kinase 2 and zinc finger protein 431 were significantly (p<0.05) higher expressed in PWG miniature pig than in $F_1$ and Duroc pigs. In conclusion, the results of the present study indicate that different proteins were expressed in different pig types, and were associated with a sperm functions and embryo development.

• Korean Journal of Veterinary Research
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• v.35 no.1
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• pp.179-185
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• 1995

The fusion rates of nuclear transplant embryos with various DC voltage were 55.6-79.2%. The significantly higher fusion rates of nuclear transplant embryos were achieved at the electric field strenght of DC 1.0-2.0kV/cm(72.0-79.2%) than DC 0.75kV/cm(55.6%, P<0.05). No significant differences in the percentage of embryos that cleaved(48.1, 55.4 and 42.6% respectively) and the percentage of cleaved embryos that developed to morulae/blastocyst(1.9, 5.3 and 1.9% respectively) could be found among the three types of in vitro culture system (Granulosa cell, BOEC co-culture and SOF, P>0.01). The age of the recipient cytoplast(30 vs 40hr) had no effect on the fusion rates and the rates of cleavage development(36.9 vs 44.1%, P>0.01).

• Parasites, Hosts and Diseases
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• v.60 no.5
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• pp.357-360
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• 2022

Excretory-secretory products (ESP) of T. vaginalis have been shown to inhibit sperm motility, viability, and functional integrity, leading to a decreased fertilization rate in vitro. This study investigated whether T. vaginalis induce apoptosis and ultrastructural changes of sperm using flow cytometry and electron microscopy. Incubation of sperm with T. vaginalis ESP increased phosphatidylserine externalization and DNA fragmentation, and decreased mitochondrial membrane potential. Transmission electron microscopy of sperm incubated with ESP revealed abnormal features such as distorted heads, broken necks, and acrosomes exocytosis. This is the first report that demonstrates a direct impact of T. vaginalis ESP on sperm apoptosis and architecture in vitro.

• Asian-Australasian Journal of Animal Sciences
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• v.14 no.10
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• pp.1360-1366
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• 2001

The aims of this study were first to evaluate the effects of different levels (20, 40 and 100%) and sources (follicular size: large, >7 mm; medium, >5-7 mm; small, 3-5 mm) of porcine follicular fluid (pFF) on the in vitro maturation (IVM) of porcine oocytes, and the effects of fertilization treatments and different culture conditions on development of fertilized oocytes were also investigated. No differences in the maturation (63.6-76.6%) and cleavage (24.8-34.3%) rates were observed among the 20,40 and 100% pFF groups (p>0.05). The cleavage rates of oocytes cultured and fertilized in 40% and 100% pFF maturation media were significantly higher than those fertilized in m199-NBCS (51.0-61.2% vs. 12.8-31.8%. p<0.05), regardless of sources of the pFF. When oocytes were fertilized in m199-NBCS followed by culture in rabbit oviducts for 4 days, the cleavage rate in 40% pFF group was better than that in 100% pFF group (46.9% vs. 32.5%, p<0.05). Two oocytes recovered from the oviducts in the 40% pFF group developed to blastocysts after IVC. However, none developed to blastocysts when fertilized in the IVM medium after being transferred to rabbit oviducts. In conclusion, addition of pFF accompanied with gonadotropins (FSH, LH) in IVM medium enhanced maturation and cleavage rates of porcine oocytes. Direct addition of sperm suspension to IVM medium may be an alternative to simplify the fertilization procedures and to reduce the mechanical lesion during manipulation. Furthermore, rabbit oviducts provide a better environment for the in vitro fertilized oocyte developing to the morula and blastocyst stages.

• Journal of Embryo Transfer
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• v.10 no.2
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• pp.171-175
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• 1995

This study was performed to establish the condition and the methods for the techniques of insertion the isolated blastomere cells into cytoplasm, in order to research the develop-mental ability of bovine embryo blastomere cells in vitro produced. After 24h in vitro ovary maturation with the ovaries from a slaughter house, in vitro fertilization was performed to the vital sperms which their mobility were decided by percoll gradient method, with 2~8 cell stage embryos, the blastomeres were isolated in $Ca^2$+. $Mg^2$+-free PBS, and following that embedded into agar and alginate solution, respectively. The rates of in vitro develop-ment are as follows ; in agar embedded 11 among 120(9.2%) 1 /2~1 /3 blastomers cleaved and 6 among 93(6.5%) 1 /4~1 /8 blastomeres cleaved. In sodium alginate-embedded 14 among 84(16.7%) 1 /2~1 /3 blastomeres cleaved and 6 among 85(7.1%) 1 /4~1 /8 blastomeres cleaved. In case of Na-alginate, the rate of the cells were better than those of agar. The results suggest that the techniques for embeeding the isolated blastomeres into gel may help cloning of bovine early embryo without nuclear transplantation.

• Korean Journal of Animal Reproduction
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• v.14 no.1
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• pp.50-56
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• 1990

Bovine oocytes obtained from follicles(2~5mm) of ovaries after slaughter were cultured in TCM 199 medium with 10~20% heat-inactivated estrus cow serum(ECS) for 25~27 hr, at 39$^{\circ}C$ under 5% CO2 in air. At the end of culture period, some oocytes were stained with 1% acetoorcein and examined for the evidence of oocyte maturation. The remainder were used to assess the potential of in vitro fertilization(IVF) with frozen-thawed spermatozoa and subsequent development in media with or without bovine oviduct epithelial cell (BOEC) co-culture. The results obtained were summarized as follows ; 1. The maturation rate of oocyte in vitro in TCM 199 medium with 15% ECS group(76.3) was superior to 10% ECS group(68.3%) and 20% ECS group(64.5%). 2. The IVF rates of oocytes matured in vitro, and formation rate of male and female pronuclei were 63.6%(77/121) and 93.5%(72/77), respectively. The incidence of polyspermy was very low(2.4%). 3. Of 73 oocytes fertilized in vitro and cultured in TCM 199 medium with 10% fetal calf serum for 7 days, 41(56.3%) were cleaved over 2-cell and only 1(2.4%) was developed beyond the 16-cell stage. 4. Of 76 oocytes co-cultured with BOEC, 58(76.3%) were cleavaged and 23(39.7%) were developed to morula and blastocyst stage. The results of this study indicate that co-culture with BOEC deserved a positive effect on the IVF oocyte development through the 16-cell block.