• Clinical and Experimental Reproductive Medicine
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• 제29권2호
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• pp.83-90
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• 2002

Objective : The purpose of the current series of experiments were to assess the effect of GM-CSF, as a medium supplement, on the development of mouse embryos and the expression of LIF and IL-1? mRNA. Materials and Methods: Mouse 2-cell embryos were collected from the oviducts of 6 weeks old ICR mice at 48 hours after hCG injection. Embryos were cultured in P-1 medium supplemented with mouse GM-CSF (0, 1, 5, 10 ng/ml). The embryo development to blastocysts and hatching blastocysts was assessed and the cell number in blastocyst was also examined. Using RT-PCR, the expressions of LIF and IL-1? mRNA in blastocyst were evaluated in the GM-CSF supplemented group and control group. Results: In mouse, the addition of GM-CSF increased the percentage of blastocysts (65.5%, 68.6%, 73.0% and 76.1% for control and 1, 5 and 10 ng/ml, respectively), and increased the proportion of hatching blastocysts (35.2%, 36.4%, 43.2% and 53.0% for control and 1, 5 and 10 ng/ml, respectively). The mean cell numbers in blastocyst were significantly increased in GM-CSF supplemented groups compared to control group. LIF and IL-1? expression in blastocyst were significantly higher in GM-CSF supplemented group than in control group. Conclusion: The results of experiment by mouse embryos showed beneficial effects of GM-CSF as a medium supplement. Furthermore, the addition of GM-CSF significantly increased the expression of LIF and IL-1? in mouse embryos. These results suggest that GM-CSF might be a important molecule in embryo implantation.

• 한국동물번식학회:학술대회논문집
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• 한국동물번식학회 2003년도 학술발표대회 발표논문초록집
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• pp.71-71
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• 2003

Granulocyte-macrophage colony-stimulating factor (GM-CSF) is a multifunctional cytokine that has been implicated in the regulation of pre-implantation embryo development across several species. The aim of this study was to determine the effects of mouse granulocyte-macrophage colony-stimulating factor (mGM-CSF) on development of porcine parthenotes and nuclear transferred embryos, and on their expression of implantation-related genes. In the presence of bovine serum albumin, mGM-CSF did not increase the percentage of oocytes that developed to the blastocyst stage and at day 7 did not increase oocyte cell number. Addition of 10 mM GM-CSF to protein-free culture medium significantly increased the compaction and blastocoel formation of 1- to 2-cell parthenotes and cloned embryos developing in vitro. However, cell number was not increased when they were cultured in the presence of GM-CSF. Semi-quantitative reverse transcripts polymerase chain reaction (RT-PCR) revealed that mGM-CSF enhances mRNA expression of the leukemia inhibitory factor receptor, but does not influence interleukin-6 or sodium/glucose co-transporter protein gene expression in blastocyst stage parthenotes. These results suggest that mGM-CSF may enhance viability of porcine embryos developing in vitro in a defined culture medium.

• Clinical and Experimental Reproductive Medicine
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• 제47권2호
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• pp.114-121
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• 2020

Objective: Despite extensive research on implantation failure, little is known about the molecular mechanisms underlying the crosstalk between the embryo and the maternal endometrium, which is critical for successful pregnancy. Profilin 1 (PFN1), which is expressed both in the embryo and in the endometrial epithelium, acts as a potent regulator of actin polymerization and the cytoskeletal network. In this study, we identified the specific role of endometrial PFN1 during embryo implantation. Methods: Morphological alterations depending on the status of PFN1 expression were assessed in PFN1-depleted or control cells grown on Matrigel-coated cover glass. Day-5 mouse embryos were cocultured with Ishikawa cells. Comparisons of the rates of F-actin formation and embryo attachment were performed by measuring the stability of the attached embryo onto PFN1-depleted or control cells. Results: Depletion of PFN1 in endometrial epithelial cells induced a significant reduction in cell-cell adhesion displaying less formation of colonies and a more circular cell shape. Mouse embryos co-cultured with PFN1-depleted cells failed to form actin cytoskeletal networks, whereas more F-actin formation in the direction of surrounding PFN1-intact endometrial epithelial cells was detected. Furthermore, significantly lower embryo attachment stability was observed in PFN1-depleted cells than in control cells. This may have been due to reduced endometrial receptivity caused by impaired actin cytoskeletal networks associated with PFN1 deficiency. Conclusion: These observations definitively demonstrate an important role of PFN1 in mediating cell-cell adhesion during the initial stage of embryo implantation and suggest a potential therapeutic target or novel biomarker for patients suffering from implantation failure.

• Clinical and Experimental Reproductive Medicine
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• 제28권3호
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• pp.235-245
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• 2001

Objectives: The purpose of this present study was to compare mouse embryo development in 3 commercial media and hatching competence of mouse embryo with or without enzymatic treatment. Methods: Collected 375 mouse embryos were divided into three groups, and then cultured in IVF-20 (G2), Medicult IVF (M3), P-1 (blastocyst M), respectively. Three day mouse morulae were cultured in G2 media treated with pronase. The results were analyzed using Chi-square test, and considered statistically significant when p<0.01. Results: The developmental rate of 2 cell mouse embryo after 72 hours was highest in IVF-20 (G2) among conventional 3 media. The hatching rate of mouse morulae was low when clultured in G2 media without pronase during 48 hours. However, it was higher when cultured in media treated with $1{\mu}g/ml$, $2.5{\mu}g/ml$, $5{\mu}g/ml$ pronase, respectively. Conclusions: Using good media and digestion of zona pellucida with enzymatic treatment improve development and hatching rate of embryo. Therefore, implantation and pregnancy rate could be improved.

• Macromolecular Research
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• 제12권2호
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• pp.219-224
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• 2004

In this work, we prepared hydrogels for wound dressing from a mixture of poly(vinyl alcohol) (PVA) and alginate using the $\^$60/Co ${\gamma}$-ray irradiation technique. We examined the physical properties of these hydrogels, including gelation, water absorptivity, and gel strength, to evaluate the applicability of these hydrogels for wound dressings. The biocompatibility of these hydrogels was also evaluated in vitro, in cultures of mouse fibroblasts, and in vivo, by subcutaneous implantation studies in rats. The gel content and strength increased upon increasing the radiation dose and upon decreasing the concentration of alginate. The degree of swelling was inversely proportional to the gel content and strength. The degree of cytotoxicity of the ${\gamma}$-ray-treated hydrogels was ca. 60% compared to the (－) control (serum) after 1 day of incubation. When the incubations were prolonged up to 2 days, the toxicity of all the samples decreased remarkably and reached that of the control. Subcutaneous implantation studies in rats indicated that foreign body reactions occurring around the implanted hydrogels were moderate and became minimal upon increasing the implantation time.

• 한국동물번식학회:학술대회논문집
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• 한국동물번식학회 2004년도 춘계학술발표대회
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• pp.225-225
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• 2004

Acyl-CoA synthetase 4(ACS4) is an arachidonate-preferring enzyme aboundant in steroidogenic tissues and postulated to modulate eicosanoid production. Most of arachidonate present in cells is esterified predominantly in phospholipids. After its release by the action of calcium-dependent phospholipases, arachidonate can be converted to prostaglandins, thromboxanes, and leukotrenes via the cyclooxygenas and lipoxygenase pathways, respectively, depending on the cell type.(omitted)

• Reproductive and Developmental Biology
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• 제30권2호
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• pp.75-79
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• 2006

The developmental regulation of the preimplantation mammalian embryos is a fundamental step for preparing the implantation and it may be regulated by several autocrine and paracrine factors including platelet-activating factor. PAF improved the embryonic survival and implantation but its role during blastocyst development is still largely unknown. In this study, the effects and the possible pathway of PAF on developmental regulation of blastocyst to hatching stage were investigated. Developmental pattern in hatching embryo was a concentration-response curve showing maximal activity at 1 nM PAF, with decreasing activity at higher concentrations. $50{\mu}M$ 1-(5-isoquinolimnesulfonyl)-2-methylpiperazinme dihydrochloride (H-7), a PKC inhibitor, inhibited the progression of blastocyst to hatching embryo. In addition H-7 blocked the PAF effects on the blastocyst development. Besides tetradecanoylphorbol acetate (TPA), a PKC activator stimulated development of blastocyst to the hatching stage. These finding revealed that PAF support the blastocyst development to the hatching embryo. Also it is suggested that PAF action pathways in hatching supporting include the PKC signaling pathway.

• Reproductive and Developmental Biology
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• 제31권1호
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• pp.21-28
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• 2007

During early embryo development, Oct-4 is an important transcription factor for the early differentiation the present study was first examined methylation status in distal enhancer and promoter region of Oct-4 during mouse pre-implantation embryo development. In oocyte and sperm, high methylation was observed in both distal and proximal of promoter in Oct-4. Following fertilization relatively high methylation level remained until 8-cell stage embryos, but decreased at the morula and blastocyst stage. Specific gene knock down of Oct-4 by siRNA injection into zygote induced higher methylation rates of both distal and proximal region of promoter of Oct-4. These results suggest a functional link between the DNA methylation status of distal and promoter resign in the Oct-4 gene and the gene sequence-specific transcriptional silencing by exogenous siRNA injection during mouse preimplantation embryos.

• 한국가축번식학회지
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• 제22권3호
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• pp.287-292
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• 1998

본 실험은 1.48$\mu\textrm{m}$ diode laser의 인간배에 대한 적용 가능성 여부를 조사하기 위한 예비실험으로, 체외생산된 생쥐배에 1.48$\mu\textrm{m}$ diode laser를 이용한 zona pellucida (ZP) drilling 처리가 배의 부화와 체내발달에 효과적인지를 조사하고자 실시하였다. 그 결과는 다음과 같다. 발달단계가 상이한(4-세포기배, 배반포기배) 생쥐배에 laser ZP drilling 처리한 후 72시간 (배반포기배) 또는 120시간 (4-세포기배) 동안 배양하였던 바, laser ZP drilling 처리를 받은 배반포기배의 부화율(81.8%)이 대조군 (54. 2%)이나 laser ZP drilling 처리를 받은 4-세포기배 (45.5%) 보다 유의하게 높게 나타났다(p<0.05). 또한, laser ZP drilling된 배반포기배를 가임신이 유도된 대리모에 이식하였던 바, 처리군 (48.7%)의 착상율이 대조군 (43.6%)보다 약간 높게 나타났다. 한편, 임신된 대리모 일부는 분만을 유도하였던 바, 태어난 모든 새끼는 처리군에 관계없이 정상적인 염색체수 (n=40), 정상적인 성장과 생식기능을 나타내었다. 이러한 결과는 1.48$\mu\textrm{m}$ diode laser를 이용한 ZP drilling이 생쥐배의 부화를 증진시키고 정상적인 임신을 유도할 수 있어 인간배에 대한 적용 간으성을 시사한다고 하겠다.

• Clinical and Experimental Reproductive Medicine
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• 제24권2호
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• pp.233-239
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• 1997

Embryos of most mammalian species grown in vitro would undergo developmental arrest at the approximate time of genomic activation. Stage-specific cell block and the resulting rapid loss of embryo viability in conventional culture media have limited the duration for which embryos may be cultured prior to transfer. As a result, embryos are usually transferred to the uterus at the 4-to 8-cell stage to avoid the loss of viability associated with long-term in vitro culture. Early transfer has led to asynchrony of the endometrium-trophectoderm interaction at the time of implantation and a resultant reduction in the rate of implantation. To overcome these problems, a variety of co-culture systems has been devised in which embryos can develop for a longer period prior to embryo transfer. Vero cells, derived from African green monkey kidney, share a common embryologic origin with cells from the genital tract. In addition, they are potentially safe to use, since they are highly controlled for viruses and other contaminants. Therefore, co-culture using Vero cells has been widely utilized to enhance embryo viability and development, although not without controversies. We thus designed a series of experiments to demonstrate whether Vero cells do indeed enhance mouse embryo development as well as to compare the efficacy of co-culturing mouse 1-cell embryos on Vero cell monolayer in both Ham's F-10 and human tubal fluid (HTF) culture media. 1-cell stage ICR mouse embryos were cultured either in the presence of Vero cells (Group A) or in conventional culture medium alone (Group B). In Ham's F-10 significantly more 3-to-8cell embryos developed in group A than group B (59.8 versus 10.0%; p<0.01). In contrast, there was no significant difference in embryonic development both group A and group B in HTF. However, significant differences were noted only in later embryonic stage (13 and 0%; p<0.05 of group A and B respectively, hatching or hatched). In Ham's F-10, we also could observe the beneficial effect of Vero cell on hatching process (70.7 and 42.1%; p<0.05 of group A and group B respectively).