ABSTRACT
OBJECTIVES: Our objective was to investigate the effect of coculture system on in vitro fertilization and development of mice embryos. METHODS: F1 hybrid mice were superovulated with PMSG/hCG. Recruited oocytes were divided into three subgroups which are control(subgroup a), Vero cell coculture(subgroup b) and human amniocyte coculture subgroup (subgroup c) respectively. For 3 subgroups, we observed fertilization after 24 hours of incubation. In vitro fertilized early 2-cell stage embryos were allocated to Group I and in vivo fertilized early 2-cell stage embryos were allocated to Group II. Also, each group was divided into control (subgroup a), Vero cell coculture(subgroup b) and human amniocyte coculture subgroup(subgroup c) respectively. For 6 subgroups, we observed in vitro development to blastocyst and that to hatching blastocyst after 120 hours of incubation. RESULTS: As to recruited oocytes, the in vitro fatilization rate of subgroup a was significantly higher than that of subgroup b and subgroup c (P<0.05). In Group I, the developmental rates were not significantly different between the three subgroups. But in Group II, the developmental rates to hatching blastocysts of subgroup b and c were significantly higher than that of subgroup a (p<0.05). CONCLUSION: The development of the in vitro fertilized mouse embryos seemed to be independent of physiologic condition which we think coculture system may give to the embryos. The independent developmental capability of the in vitro fertilized embryos might be obtained through a certain intracellular mechanism for which there should be the need of many more investigations to be verified.
Subject(s)
Animals , Humans , Mice , Blastocyst , Coculture Techniques , Embryonic Structures , Fertilization , Fertilization in Vitro , Oocytes , Vero CellsABSTRACT
BACKGROUND: Epidermal growth factor receptor (EGFR) is overexpressed in the tissue of various malignancies including carcinoma of the breast, lung, esophagus, cervix, and ovary. In patients with cervical neoplasia, there may be a relationship between the expressions of EGFR in cervical neoplastic tissue and serum. METHODS: The expression of EGFR was determined in cervical tissues from 23 cervical intraepithelial neoplasia (CIN) patients and 16 invasive cervical carcinoma patients using immunohistochemical staining and the level of serum EGFR ECD (extracellular domain) was measured in serum from 17 CIN patients and 14 cervical carcinoma patients using ELISA (enzyme-linked immunosorbent assay). RESULTS: The expression of EGFR in cervical tissue was significantly increased as normal cervical tissue progressed to CIN then to invasive cervical carcinoma (p=0.009). And the mean level of serum EGFR according to the histologic diagnosis of normal cervix, CIN, invasive cervical carcinoma was 23.18+/-1.92 fmol/ml, 23.49+/-8.95 fmol/ml, and 30.46+/-19.72 fmol/ml, respectively. The mean level of serum EGFR was higher in invasive cervical carcinoma than that of normal cervix or CIN. But there was no significant statistical difference (p=0.471). Also the mean level of serum EGFR according to the intensity of immunohistochemical staining in negative (-), weakly positive (+), positive (++), and strongly positive (+++) staining was 19.36+/- 3.12 fmol/ml, 20.99+/-3.59 fmol/ml, 29.08+/-16.86 fmol/ml, and 24.34+/-10.35 fmol/ml, respectively. The mean level of serum EGFR in positive (++) and strongly positive (+++) staining was higher than in negative (-) staining, but there was no significant statistical difference (p=0.450). CONCLUSIONS: The authors believe that the expression of EGFR in cervical neoplastic tissue could be used as a marker for reflecting the malignant transformation of cervical epithelial cells. Although the mean level of serum EGFR in invasive cervical carcinoma was higher than in normal cervix and CIN, and the mean level of serum EGFR in positive (++) and strongly positive (+++) immunohistochemical staining was higher than in negative (-) staining, there was no significant statistical difference, possibly due to the limited number of cases in this preliminary study. So, the authors believe that the level of serum EGFR may have a similar role as a tumor marker like the EGFR expression in cervical neoplastic tissue. This study should be continued further with more cases and the relationship between the level of serum EGFR and prognostic parameters of uterine cervical carcinoma need to be analyzed.
Subject(s)
Female , Humans , Breast , Uterine Cervical Dysplasia , Cervix Uteri , Diagnosis , Enzyme-Linked Immunosorbent Assay , Epithelial Cells , Esophagus , Lung , Ovary , ErbB ReceptorsABSTRACT
OBJECT: This study was carried out to investigate the effect of pentoxifylline on in vitro fertillization and developmen of preimplantation stage of mouse embryos. MATERIAL AND METHODS:F1 hybrid mice was superovulated with PMSG/hCG and mouse oocytes were recruited. After the normal sperms were incubated with PTX before in vitro fertilization, it was observed whether the fertilization and embryo development was affected or not by the sperm preparation(washing, dilution and no washing or no dilution). And after 1-cell and 2-cell stage of mouse embryos were incubated with PTX, the development to hatching blastocyst was also observed. RESULTS: When in vitro fertilization was revealed by using the washed normal sperms after 0, 3.6 and 7.2 mM PTX incubation, the fertilization rates were 92.5%, 48.8%, 36.8%, respectively. So 3.6 and 7.2 mM groups presented significantly low fertilizatin rate, but the development rates were 93.9%, 85.0%, 95.2%, respectively. Therefore, there were no significant difference between each group. When in vitro fertilization was revealed by using the diluted normal sperms after 0, 3.6, and 7.2 mM PTX incubation, the fertilization rates were 58.6%, 5.4%, 9.4%, respectively. So 3.6 and 7.2 mM groups presented significantly low fertilization rate. The developmental rates were 88.2%, 100%, 100%. And there were no significant difference between each group. When in vitro fertilization was revealed by using the not washed and not diluted normal sperms after 0, 3.6 and 7.2 mM PTX incubation, the fertilizatin rates were 61.2%, 5.7%, 3.8%, respectively. 3.6 and 7.2 mM group presented significantly low fertilization rate. The development rates were 73.3%, 0%, 0%, respectively. So 3.6, 7.2 mM group presented significantly low developmental rate. After 1-cell stage of mouse embryos were incubated in 0, 5, 10, 50 nM of PTX, the development rates were not significantly different among them. After 2-cell stage of mouse embryos were incubated in 0, 5, 10, 50 nM of PTX, the development rates were not significantly different among them. CONCLUSION: In conclusion, when PTX is used in in vitro fertilization program with normal sperms, it may affect the fertilization and embryo development in high concentration. And if PTX concentration is very low, the developmental rate would not be affected. So PTX must not be used to normal sperms and where use of PTX is indicated, it is recommended that remainder PTX must be removed as completely as possible.
Subject(s)
Animals , Female , Mice , Pregnancy , Blastocyst , Embryonic Development , Embryonic Structures , Fertilization , Fertilization in Vitro , Oocytes , Pentoxifylline , SpermatozoaABSTRACT
No abstract available.
Subject(s)
Animals , Humans , Mice , Embryonic Structures , Fertilization in Vitro , Fetal Blood , Quality Control , Sperm Motility , SpermatozoaABSTRACT
No abstract available.
ABSTRACT
No abstract available.