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Revista Brasileira de Ginecologia e Obstetrícia. 2021;43(11):878-882
, , , , , , , , Non-invasive preimplantation genetic testing for aneuploidies (niPGT-A) aiming to assess cell-free embryonic DNA in spent culturemedia is promising, especially because it might overcome the diminished rates of implantation caused by the inadequate performance of trophectoderm (TE) biopsy. Our center is part of the largest study to date assessing the concordance between conventional PGT-A and niPGT-A, and we report here the delivery of the first baby born in Brazil using niPGT-A. The parents of the baby were admitted to our center in 2018. They did not present history of infertility, and they were interested in using in vitro fertilization (IVF) and PGT-A in order to avoid congenital anomalies in the offspring. A total of 11 (3 day-5 and 8 day-6) expanded blastocysts were biopsied, and the spent culture media (culture from day-4 to day-6) from 8 day-6 blastocysts were collected for niPGT-A. Overall, 7 embryos yielded informative results for trophectoderm (TE) and media samples. Among the embryos with informative results, 5 presented concordant diagnosis between conventional PGTA and niPGT-A, and 2 presented discordant diagnosis (1 false-positive and one falsenegative). The Blastocyst 4, diagnosed as 46, XY by both niPGT-A and conventional PGTA, was warmed up and transferred, resulting in the birth of a healthy 3.8 kg boy in February 2020. Based on our results and the recent literature, we believe that the safest current application of niPGT-A would be as a method of embryo selection for patients without an indication for conventional PGT-A. The approximate 80% of reliability of niPGT-A in the diagnosis of ploidy is superior to predictions provided by other noninvasive approaches like morphology and morphokinetics selection.
Summary
Revista Brasileira de Ginecologia e Obstetrícia. 2021;43(11):878-882
, , , , , , , , Non-invasive preimplantation genetic testing for aneuploidies (niPGT-A) aiming to assess cell-free embryonic DNA in spent culturemedia is promising, especially because it might overcome the diminished rates of implantation caused by the inadequate performance of trophectoderm (TE) biopsy. Our center is part of the largest study to date assessing the concordance between conventional PGT-A and niPGT-A, and we report here the delivery of the first baby born in Brazil using niPGT-A. The parents of the baby were admitted to our center in 2018. They did not present history of infertility, and they were interested in using in vitro fertilization (IVF) and PGT-A in order to avoid congenital anomalies in the offspring. A total of 11 (3 day-5 and 8 day-6) expanded blastocysts were biopsied, and the spent culture media (culture from day-4 to day-6) from 8 day-6 blastocysts were collected for niPGT-A. Overall, 7 embryos yielded informative results for trophectoderm (TE) and media samples. Among the embryos with informative results, 5 presented concordant diagnosis between conventional PGTA and niPGT-A, and 2 presented discordant diagnosis (1 false-positive and one falsenegative). The Blastocyst 4, diagnosed as 46, XY by both niPGT-A and conventional PGTA, was warmed up and transferred, resulting in the birth of a healthy 3.8 kg boy in February 2020. Based on our results and the recent literature, we believe that the safest current application of niPGT-A would be as a method of embryo selection for patients without an indication for conventional PGT-A. The approximate 80% of reliability of niPGT-A in the diagnosis of ploidy is superior to predictions provided by other noninvasive approaches like morphology and morphokinetics selection.
Summary
Revista Brasileira de Ginecologia e Obstetrícia. 2018;40(12):763-770
The aim of the present study was to provide a better understanding of the specific action of two follicle-stimulating hormone (FSH) isoforms (β-follitropin and sheep FSH) on the membrane potential of human cumulus cells.
Electrophysiological data were associated with the characteristics of the patient, such as age and cause of infertility. The membrane potential of cumulus cells was recorded with borosilicate microelectrodes filled with KCl (3 M) with tip resistance of 15 to 25 MΩ. Sheep FSH and β-follitropin were topically administered onto the cells after stabilization of the resting potential for at least 5 minutes.
In cumulus cells, the mean resting membrane potential was - 34.02 ± 2.04 mV (n = 14). The mean membrane resistance was 16.5 ± 1.8 MΩ (n = 14). Sheep FSH (4 mUI/mL) and β-follitropin (4 mUI/mL) produced depolarization in the membrane potential 180 and 120 seconds after the administration of the hormone, respectively.
Both FSH isoforms induced similar depolarization patterns, but β-follitropin presented a faster response. A better understanding of the differences of the effects of FSH isoforms on cell membrane potential shall contribute to improve the use of gonadotrophins in fertility treatments.
Summary
Revista Brasileira de Ginecologia e Obstetrícia. 2018;40(12):763-770
The aim of the present study was to provide a better understanding of the specific action of two follicle-stimulating hormone (FSH) isoforms (β-follitropin and sheep FSH) on the membrane potential of human cumulus cells.
Electrophysiological data were associated with the characteristics of the patient, such as age and cause of infertility. The membrane potential of cumulus cells was recorded with borosilicate microelectrodes filled with KCl (3 M) with tip resistance of 15 to 25 MΩ. Sheep FSH and β-follitropin were topically administered onto the cells after stabilization of the resting potential for at least 5 minutes.
In cumulus cells, the mean resting membrane potential was - 34.02 ± 2.04 mV (n = 14). The mean membrane resistance was 16.5 ± 1.8 MΩ (n = 14). Sheep FSH (4 mUI/mL) and β-follitropin (4 mUI/mL) produced depolarization in the membrane potential 180 and 120 seconds after the administration of the hormone, respectively.
Both FSH isoforms induced similar depolarization patterns, but β-follitropin presented a faster response. A better understanding of the differences of the effects of FSH isoforms on cell membrane potential shall contribute to improve the use of gonadotrophins in fertility treatments.
Summary
Revista Brasileira de Ginecologia e Obstetrícia. 2009;31(3):138-141
DOI 10.1590/S0100-72032009000300006
PURPOSE: to report three cases of spontaneous gestation in women with polycystic ovarian syndrome (PCOS), that occurred in the months subsequent to transvaginal oocyte retrieval for in vitro maturation (IVM). METHODS: three infertile patients with PCOS, submitted to oocytes' IVM without previous ovarian stimulation, were included in the study. During the procedure of oocytes' collection, each ovary was drilled from four to eight times. RESULTS: none of the patients got pregnant with the IVM technique. Evaluating the cases' follow-up, in seven months after the procedure, the three patients got pregnant without the help of techniques of assisted reproduction, which resulted in three births. CONCLUSIONS: the multiple drillings in the ovary of these patients with PCOS, during the process to collect oocytes, may have contributed to their pregnancy in the months following the procedure.
Summary
Revista Brasileira de Ginecologia e Obstetrícia. 2009;31(3):138-141
DOI 10.1590/S0100-72032009000300006
PURPOSE: to report three cases of spontaneous gestation in women with polycystic ovarian syndrome (PCOS), that occurred in the months subsequent to transvaginal oocyte retrieval for in vitro maturation (IVM). METHODS: three infertile patients with PCOS, submitted to oocytes' IVM without previous ovarian stimulation, were included in the study. During the procedure of oocytes' collection, each ovary was drilled from four to eight times. RESULTS: none of the patients got pregnant with the IVM technique. Evaluating the cases' follow-up, in seven months after the procedure, the three patients got pregnant without the help of techniques of assisted reproduction, which resulted in three births. CONCLUSIONS: the multiple drillings in the ovary of these patients with PCOS, during the process to collect oocytes, may have contributed to their pregnancy in the months following the procedure.
Summary
Revista Brasileira de Ginecologia e Obstetrícia. 2003;25(10):711-715
DOI 10.1590/S0100-72032003001000003
PURPOSE: to compare the characteristics of human semen preserved at +4ºC and at -196ºC for 24 h and to determine which technique is indicated for use in specific procedures. METHODS: semen samples of 24 voluntaries were analyzed after collection and divided into two aliquots, one of them cooled (+4ºC) and the other frozen (-196ºC). Samples were kept at low temperatures for 24 h and then at room temperature for 30 minutes (T1), capacitated (T2) and kept at +37ºC for 90 minutes (T3), being analyzed regarding count and progressive motility at T1, T2 and T3. The General Linear Model was used to analyze results obtained with different techniques, while Wilcoxon's test was used to compare results obtained in two different moments using the same technique (a = 5% e p<0,05). RESULTS: data were missed in one fresh semen sample, in one sample after preservation, in five samples after capacitation and in two samples after incubation. The average number of total motile sperm/mL (NTMS) in fresh semen was 39.7 million (1.3-104.0). After preservation, the average NTMS in cooled semen was 9.6 million (0-37.4) and in frozen semen 8.7 million (0-41.2). After capacitation, the average NTMS was 5.4 million either in cooled (0-21.7), or in frozen semen (0-28). After incubation, the average NTMS in cooled semen was 9.8 million (0-40.5) and in frozen 4.4 million (0-25.6). Concerning count, progressive motility and NTMS, there was no significant difference (p>0,05) between techniques in the three moments of observation. In cooled samples, there was no difference between variables after capacitation and after incubation, but, in frozen semen, count was significantly greater after capacitation. CONCLUSIONS: although there has been no significant difference between semen count and progressive motility in both techniques, the use of cooled semen is recommended for specific procedures within a short time period due to its simplicity and low cost. When frozen semen is necessary, we recommend its use soon after capacitation in order to avoid loss in quality.
Summary
Revista Brasileira de Ginecologia e Obstetrícia. 2003;25(10):711-715
DOI 10.1590/S0100-72032003001000003
PURPOSE: to compare the characteristics of human semen preserved at +4ºC and at -196ºC for 24 h and to determine which technique is indicated for use in specific procedures. METHODS: semen samples of 24 voluntaries were analyzed after collection and divided into two aliquots, one of them cooled (+4ºC) and the other frozen (-196ºC). Samples were kept at low temperatures for 24 h and then at room temperature for 30 minutes (T1), capacitated (T2) and kept at +37ºC for 90 minutes (T3), being analyzed regarding count and progressive motility at T1, T2 and T3. The General Linear Model was used to analyze results obtained with different techniques, while Wilcoxon's test was used to compare results obtained in two different moments using the same technique (a = 5% e p<0,05). RESULTS: data were missed in one fresh semen sample, in one sample after preservation, in five samples after capacitation and in two samples after incubation. The average number of total motile sperm/mL (NTMS) in fresh semen was 39.7 million (1.3-104.0). After preservation, the average NTMS in cooled semen was 9.6 million (0-37.4) and in frozen semen 8.7 million (0-41.2). After capacitation, the average NTMS was 5.4 million either in cooled (0-21.7), or in frozen semen (0-28). After incubation, the average NTMS in cooled semen was 9.8 million (0-40.5) and in frozen 4.4 million (0-25.6). Concerning count, progressive motility and NTMS, there was no significant difference (p>0,05) between techniques in the three moments of observation. In cooled samples, there was no difference between variables after capacitation and after incubation, but, in frozen semen, count was significantly greater after capacitation. CONCLUSIONS: although there has been no significant difference between semen count and progressive motility in both techniques, the use of cooled semen is recommended for specific procedures within a short time period due to its simplicity and low cost. When frozen semen is necessary, we recommend its use soon after capacitation in order to avoid loss in quality.
Summary
Revista Brasileira de Ginecologia e Obstetrícia. 2003;25(8):553-559
DOI 10.1590/S0100-72032003000800003
PURPOSE: to verify vitrification techniques using 6 M DMSO to cryopreserve in vitro matured bovine oocytes, and to assess the effects of the time of exposure to vitrification solutions (VS). METHODS: dilutions of VS were prepared from the stock VS (VS 100%) consisting of 6 M DMSO to give 25 and 65% DMSO solutions. Bovine oocytes were in vitro matured for 18-22 h. Matured oocytes were placed first into 25% VS, at room temperature for 5 min, then transferred to 65% VS, before being pipetted into the 100% VS in plastic straws. Three experimental groups were formed: in the first group, time of pipetting through 65% VS and loading the straw took up to 60 s, in the second group it did not exceed 30 s. For thawing, straws were held in air for 10 s and then in a water bath for 10 s. The contents of each straw were expelled in sucrose solution and held for 5 min. In the third experimental group, oocytes went through all VS, but were not vitrified. All retrieved oocytes were inseminated. For control, fresh, in vitro matured oocytes were inseminated. RESULTS: after vitrification, 69.1 and 59.8% of the oocytes were retrieved from the 30 s and 60 s groups, respectively, and 93 and 89% of these oocytes appeared morphologically normal 24 h after insemination, respectively. In the group of oocytes exposed without vitrification, 75.6% were retrieved and 84.7% were morphologically viable, 24 h after insemination. No fertilization was observed in the experimental groups. Among controls, 65.4% were fertilized. CONCLUSIONS: the vitrification technique using 6 M DMSO is not a feasible approach to cryopreserve in vitro matured bovine oocytes. Decreasing the time of exposure to VS did not overcome deleterious effects of the procedure on the fertilizability of oocytes. Improvements in the technique are needed to protect the zona pellucida and oolemma.
Summary
Revista Brasileira de Ginecologia e Obstetrícia. 2003;25(8):553-559
DOI 10.1590/S0100-72032003000800003
PURPOSE: to verify vitrification techniques using 6 M DMSO to cryopreserve in vitro matured bovine oocytes, and to assess the effects of the time of exposure to vitrification solutions (VS). METHODS: dilutions of VS were prepared from the stock VS (VS 100%) consisting of 6 M DMSO to give 25 and 65% DMSO solutions. Bovine oocytes were in vitro matured for 18-22 h. Matured oocytes were placed first into 25% VS, at room temperature for 5 min, then transferred to 65% VS, before being pipetted into the 100% VS in plastic straws. Three experimental groups were formed: in the first group, time of pipetting through 65% VS and loading the straw took up to 60 s, in the second group it did not exceed 30 s. For thawing, straws were held in air for 10 s and then in a water bath for 10 s. The contents of each straw were expelled in sucrose solution and held for 5 min. In the third experimental group, oocytes went through all VS, but were not vitrified. All retrieved oocytes were inseminated. For control, fresh, in vitro matured oocytes were inseminated. RESULTS: after vitrification, 69.1 and 59.8% of the oocytes were retrieved from the 30 s and 60 s groups, respectively, and 93 and 89% of these oocytes appeared morphologically normal 24 h after insemination, respectively. In the group of oocytes exposed without vitrification, 75.6% were retrieved and 84.7% were morphologically viable, 24 h after insemination. No fertilization was observed in the experimental groups. Among controls, 65.4% were fertilized. CONCLUSIONS: the vitrification technique using 6 M DMSO is not a feasible approach to cryopreserve in vitro matured bovine oocytes. Decreasing the time of exposure to VS did not overcome deleterious effects of the procedure on the fertilizability of oocytes. Improvements in the technique are needed to protect the zona pellucida and oolemma.