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Site of gonadotrophin injection affects the quality of superovulated ova
161 INITIAL ~ T S
SITE OF GONADOTROPHIN INJECTION AFFECTS THE QUALITY OF SUPEROVULATED OVA. R. Kankondl and E. Lehtonen. Department of Pathology, Unlver sl ty of Hel s I nkl, Haartmaninkatu 3, SF-00290 Helslnkl, Finland.
OF AN (~T-PATIE~ IVF-PIK)(E~ INVOLV]~
BOYH OOCYTE A~D EMERYO ~ . M. Saaranan, S. Vilska~ M. Vierula, S. Saarikoski and T. Vanha-Perttula. Departments of Armtomy and Obstetrics and Gynecology, University of Kuopio, 70211 Kuopio, Finland. An out-patient IVF-progrem was started in September 1986 in collaboration with the Clinic of Obstetrics and Gynecology of the University Hospital and the Anatomy Department, which are about i km apart and the intervening transport time is about 15 rain. Superovulatien ir~h~ctiou ~ms performed according to individually planned schemes. The aspiration of follicles was carried out using transvaginal probe. The follicular fluids were transported in a thermo6tated box at 37 °C from the Clinic to the Anatomy Department. The oocytes were inseminated 4-6 h after recovery in HAM F-10 medium supplemented with 10-20 % patient serum. The embryos were loaded to the W~11~ce catheter and then transported in a glass tube filled with pre-eq,1111brated i00 % serum in the pocket to the Clinic, where the embryo transfer was performed as an out-patient procedure. This study consisted of 34 consecutive treatment cycles within 4 mounths. The mean age of the women was 31.8 (26 to 40) years. Of these cycles, 29 showed good ovarian st1,~;l~tion and 87 oocytes (mean = 3.0/puncture) were recovered. The fertilization rate was 51%. Total number of embryos in 21 transfers was 44 (2.1/ transfer) and three pregnancies were obtained, 2 subclinical and i with ultrasound verified pregnancy going on normally. It seems that human oocytes and e~bry~ can be transported safely in suitable conditions between the laboratory and the out-patient clinic.
Intraperltoneal (l.p.) injection of gonadotrophlns Is widely used for superovulating mice. We investigated how the route of hormone administration affects the normality of oocytes and preimplantat lon embryos. Subcutaneous (s.c.) and l.p. injections of gonadotrophlns both caused about threefold increase in the number of ova. At 20-22 h after hCG, the proportion of abnormal oocytes in mice injected s.c. was 7.8%, and i.p., 15.4%. At 72-74 h after hCG, the corresponding proportions of abnormal ova were 18% and 22.9%. The mechanisms by which the l.p. route of hormone treatment increases the incidence of abnormallty are not known. The results suggest that s.c. rather than i.p. administration of gonadotrophlns should be used for superovulat lon of mice.
162
STUDIES ON GENOTOXIC EFFECTS OF SUPEROVULATION IN PREIMPLANTATION MOUSE EMBRYOS USING THE SISTER CHROMATID EXCHANGE (SCE) ASSAY. R.Vogel r T.Lankes~ H.Rux and H.Spielmann Bundesgesundheitsamt, D-IO00 Berlin 33 Sexual hormones are used quite commonly in therapy of human infertility and also in animal breeding. Reports about genotoxic e ~ fects of hormone induced ovulation (supero~ ulation) are controversial. Recently, some evidence was found in pre- and postimplantation embryos for effects of superovulation with human gonadotropins, since the frequem cies of sister chromatid exchanges (SCE) were increased after superovulation. We tried to confirm these results in four different mouse strains. Females were super ovulated with 5 IU PMSG and 5 IU HCG 48 hrs later. To visualise SCE, 4 and 8 celled embros as well as morulae and blastocystswere cultured in presence of BrdU for 26 hrs. After spontaneous ovulation SCE levels in embryos were very high (25-40 SCE/metaphase) compared to adult tissues like bone marrow of the mother animals (6 SCE/metaphase). SCE frequencies, however, did not change in any strain after superovulation, although it was increased significantly after exposure to genotoxic agents in vivo (Vogel and Spielmann, Tox.Lett.1986). Thus, the use of g o n m dotropins seems not to be genotoxic. 58S
SITE OF GONADOTROPHIN INJECTION AFFECTS THE QUALITY OF SUPEROVULATED OVA. R. Kankondl and E. Lehtonen. Department of Pathology, Unlver sl ty of Hel s I nkl, Haartmaninkatu 3, SF-00290 Helslnkl, Finland.
OF AN (~T-PATIE~ IVF-PIK)(E~ INVOLV]~
BOYH OOCYTE A~D EMERYO ~ . M. Saaranan, S. Vilska~ M. Vierula, S. Saarikoski and T. Vanha-Perttula. Departments of Armtomy and Obstetrics and Gynecology, University of Kuopio, 70211 Kuopio, Finland. An out-patient IVF-progrem was started in September 1986 in collaboration with the Clinic of Obstetrics and Gynecology of the University Hospital and the Anatomy Department, which are about i km apart and the intervening transport time is about 15 rain. Superovulatien ir~h~ctiou ~ms performed according to individually planned schemes. The aspiration of follicles was carried out using transvaginal probe. The follicular fluids were transported in a thermo6tated box at 37 °C from the Clinic to the Anatomy Department. The oocytes were inseminated 4-6 h after recovery in HAM F-10 medium supplemented with 10-20 % patient serum. The embryos were loaded to the W~11~ce catheter and then transported in a glass tube filled with pre-eq,1111brated i00 % serum in the pocket to the Clinic, where the embryo transfer was performed as an out-patient procedure. This study consisted of 34 consecutive treatment cycles within 4 mounths. The mean age of the women was 31.8 (26 to 40) years. Of these cycles, 29 showed good ovarian st1,~;l~tion and 87 oocytes (mean = 3.0/puncture) were recovered. The fertilization rate was 51%. Total number of embryos in 21 transfers was 44 (2.1/ transfer) and three pregnancies were obtained, 2 subclinical and i with ultrasound verified pregnancy going on normally. It seems that human oocytes and e~bry~ can be transported safely in suitable conditions between the laboratory and the out-patient clinic.
Intraperltoneal (l.p.) injection of gonadotrophlns Is widely used for superovulating mice. We investigated how the route of hormone administration affects the normality of oocytes and preimplantat lon embryos. Subcutaneous (s.c.) and l.p. injections of gonadotrophlns both caused about threefold increase in the number of ova. At 20-22 h after hCG, the proportion of abnormal oocytes in mice injected s.c. was 7.8%, and i.p., 15.4%. At 72-74 h after hCG, the corresponding proportions of abnormal ova were 18% and 22.9%. The mechanisms by which the l.p. route of hormone treatment increases the incidence of abnormallty are not known. The results suggest that s.c. rather than i.p. administration of gonadotrophlns should be used for superovulat lon of mice.
162
STUDIES ON GENOTOXIC EFFECTS OF SUPEROVULATION IN PREIMPLANTATION MOUSE EMBRYOS USING THE SISTER CHROMATID EXCHANGE (SCE) ASSAY. R.Vogel r T.Lankes~ H.Rux and H.Spielmann Bundesgesundheitsamt, D-IO00 Berlin 33 Sexual hormones are used quite commonly in therapy of human infertility and also in animal breeding. Reports about genotoxic e ~ fects of hormone induced ovulation (supero~ ulation) are controversial. Recently, some evidence was found in pre- and postimplantation embryos for effects of superovulation with human gonadotropins, since the frequem cies of sister chromatid exchanges (SCE) were increased after superovulation. We tried to confirm these results in four different mouse strains. Females were super ovulated with 5 IU PMSG and 5 IU HCG 48 hrs later. To visualise SCE, 4 and 8 celled embros as well as morulae and blastocystswere cultured in presence of BrdU for 26 hrs. After spontaneous ovulation SCE levels in embryos were very high (25-40 SCE/metaphase) compared to adult tissues like bone marrow of the mother animals (6 SCE/metaphase). SCE frequencies, however, did not change in any strain after superovulation, although it was increased significantly after exposure to genotoxic agents in vivo (Vogel and Spielmann, Tox.Lett.1986). Thus, the use of g o n m dotropins seems not to be genotoxic. 58S