RESUMO
As assisted reproductive technology (ART) expanded globally, several countries introduced prescribed requirements for treatment and monitoring of outcomes, as well as a licensing or accreditation requirement. While it is common for ART laboratories to be required to have an effective quality control system, the remainder of the clinic is often under less stringent regulation. Furthermore, when treatment conditions are prescribed, the standards tend to be conservative and clinics may choose to establish their own standards. Total quality management systems are now being used by an increasing number of ART clinics. In Australia and New Zealand, it is now a requirement to have a quality management system in order to be accredited and to help meet customer demand for improved delivery of ART services in these two countries.
Assuntos
Acreditação , Garantia da Qualidade dos Cuidados de Saúde , Técnicas de Reprodução Assistida/normas , Austrália , Feminino , Humanos , GravidezRESUMO
The development of one-cell mouse zygotes to the blastocyst stage in vitro has been used as a quality control for the media and handling procedures employed for human in vitro fertilization and embryo transfer (IVF/ET). One-cell mouse zygotes were placed in culture in medium containing bovine serum albumin. Aliquots of the same batch of medium containing female patients' homologous serum were used for the fertilization and culture of human oocytes. The following procedures were associated with high rates of mouse embryo development and human pregnancies following IVF/ET: adequate gassing and equilibration of the medium, double-rinsing of pipets and catheters used to handle embryos, use of a HEPES-buffered medium for manipulating embryos in the absence of an atmosphere containing 5% CO2, control of excessive temperature in the vicinity of the embryos, and ET using medium containing 50% patient's serum. The institution of these procedures gave more consistent pregnancy rates. However, there was no obvious association between fertilization and cleavage of human oocytes and the quality of the medium ascertained by the mouse embryo development test. In a continuing trial, we are comparing two culture media (modified Tyrode's and a medium formulated on the composition of human fallopian tube fluid [HTF]) and two culture techniques (culture in medium under oil in petri dishes and in loosely capped tubes). Significantly more mouse zygotes developed in HTF medium compared to Tyrode's medium. In a randomized 2 X 2 factorial trial with human IVF/ET, the highest pregnancy rate occurred when fertilization and culture were carried out in HTF medium under oil, but numbers are not yet sufficient to show any statistical difference between treatments.
Assuntos
Transferência Embrionária , Fertilização in vitro/métodos , Animais , Meios de Cultura , Técnicas de Cultura/métodos , Feminino , Humanos , Inalação , Masculino , Camundongos , Camundongos Endogâmicos , Oócitos/citologia , Oócitos/fisiologia , GravidezRESUMO
In a prospective clinical trial the pregnancy rate in patients matched for infertility status, degree of hyperstimulation, and number of oocytes recovered was unaffected by whether embryos were transferred while still pronuclear (day 1) or after they had undergone cleavage (day 2). The pregnancy rates per transfer were 27% and 22%, respectively, for the two transfer times. Unlike results of a previous study, no difference was detected in the outcome of pregnancies from the two groups.
Assuntos
Transferência Embrionária , Embrião de Mamíferos/citologia , Fertilização in vitro , Resultado da Gravidez , Divisão Celular , Ensaios Clínicos como Assunto , Estradiol/sangue , Feminino , Humanos , Indução da Ovulação , Gravidez , Progesterona/sangue , Estudos ProspectivosRESUMO
Significantly more mouse zygotes developed to blastocysts in culture in a medium formulated on the composition of human tubal fluid (HTF) than in modified Tyrode's medium (T6). In a randomized 2 X 2 factorial trial of human in vitro fertilization that compared the two media and culture under oil versus culture in loosely capped tubes, significantly more clinical pregnancies (30% of 60 transfers) were obtained with HTF medium than with T6 medium (11% of 53 transfers). Decreasing the K+ content of HTF medium to that present in T6 medium significantly decreased the number of mouse zygotes that developed in culture. Modifying Ca++ levels had no effect. It is therefore likely that the higher K+ content in HTF medium is primarily responsible for the superiority of HTF medium over T6 medium, but other differences in the composition of the two media could contribute to the results observed.
Assuntos
Líquidos Corporais , Meios de Cultura , Fertilização in vitro/métodos , Animais , Blastocisto , Ensaios Clínicos como Assunto , Transferência Embrionária , Tubas Uterinas , Feminino , Humanos , Camundongos , Oócitos/crescimento & desenvolvimento , Distribuição AleatóriaRESUMO
We measured chemotactic activity in 238 follicular fluids (FF) aspirated from 45 women who had undergone ovarian stimulation with a combination of clomiphene citrate and human menopausal gonadotropin for oocyte retrieval, in vitro fertilization, and embryo transfer. Fifteen of the treatment cycles resulted in pregnancy. The mean chemotactic activity, measured as the distance in microns granulocytic leukocytes migrated through a 3.0-micron membrane, was significantly higher in FF from conceptual cycles, compared with nonconceptual cycles. Serum chemotactic activity was significantly lower in conceptual cycles, compared with nonconceptual cycles. A chemotactic gradient appears to exist between the peripheral circulation and the ovarian follicle. The gradient favors the follicle in conceptual cycles, as indicated by the chemotactic quotient (the ratio of chemotactic activity of FF to serum). In conceptual cycles the chemotactic quotient was 1.7 +/- 0.17, compared with 0.7 +/- 0.03 for nonconceptual cycles. The presence of leukocyte chemotactic factor in FF appears to discriminate prospectively with a 90% degree of confidence between conceptual and nonconceptual in vitro fertilization and embryo transfer cycles.
Assuntos
Líquidos Corporais/fisiologia , Quimiotaxia de Leucócito , Granulócitos , Folículo Ovariano/fisiologia , Gravidez , Androstenodiona/análise , Androstenodiona/sangue , Líquidos Corporais/análise , Transferência Embrionária , Estradiol/análise , Estradiol/sangue , Feminino , Fertilização in vitro , Humanos , Progesterona/análise , Progesterona/sangueRESUMO
The development of 1-cell mouse zygotes to the blastocyst stage in vitro has been used as a quality control for the media employed for human in vitro fertilization and embryo transfer (IVF-ET). The following procedures were associated with high rates of mouse embryo development and human pregnancies following IVF-ET: adequate gassing and equilibration of the medium, double rinsing of pipettes and catheters used to handle embryos, use of a HEPES-buffered medium for manipulating embryos in the absence of an atmosphere containing 5% CO2, control of excessive temperature in the vicinity of the embryos, and ET using medium containing 50% patient's serum. The institution of these procedures gave more consistent pregnancy rates. However, there was no obvious association between fertilization and cleavage of human oocytes and the quality of the medium ascertained by the mouse embryo development test.
Assuntos
Transferência Embrionária/métodos , Fertilização in vitro/métodos , Animais , Meios de Cultura , Feminino , Humanos , Masculino , Camundongos , Controle de Qualidade , ZigotoRESUMO
A retrospective analysis was performed of 368 normally cycling women treated with a single cycle of a standard ovarian hyperstimulation regime (CC 100 mg days 5 to 9 and hMG 150 IU days 6, 8, and 10) associated with either an IVF or GIFT program. Neither the peak serum E2 level attained nor the number of days of stimulation required bore a relationship to the BMI or the total body weight of these women. Whereas the mean number of oocytes aspirated from women with BMI less than 19.1 was higher (6.4 +/- 3.2) compared with obese women (BMI greater than 27.6, 4.8 +/- 2.6), the rate of fertilization was not different for both BMI extremes. It is concluded that factors other than BMI or total body weight have more important influences on the response to hyperstimulation in normal women.
Assuntos
Índice de Massa Corporal , Peso Corporal , Indução da Ovulação , Clomifeno/uso terapêutico , Estradiol/sangue , Feminino , Humanos , Menotropinas/uso terapêutico , Estudos RetrospectivosRESUMO
STUDY OBJECTIVE: The efficacy of intrauterine insemination (IUI) of selected motile sperm. DESIGN: Prospective randomized sequential alternating cycle trial comparing IUI with luteinizing hormone (LH)-timed intercourse. SETTING: Clinical infertility service. PATIENTS: Couples selected included unexplained infertility (n = 73), cervical mucus hostility (n = 24), moderate semen defect (n = 110), and severe semen defect (n = 78). Two hundred eighty-five couples undertook 600 IUI cycles and 505 LH-timed intercourse. RESULTS: Overall, IUI was slightly more effective than LH-timed intercourse with a pregnancy rate of 6.2% versus 3.4% per cycle. When individual categories were considered only, IUI for severe semen defect was significantly better (5.6% versus 1.3%, P less than 0.05). The first IUI cycle was more effective when compared with both subsequent IUI cycles and the initial LH-timed cycle. Overall, 74% (27/37) of IUI pregnancies occurred in the first cycle. CONCLUSIONS: Compared with LH-timed intercourse, IUI provided little or no improved expectation of pregnancy but was beneficial in couples with severe semen defect. The occurrence of pregnancy was limited per cycle and confined essentially to the initial cycle of treatment. Continued IUI is considered to be unrewarding.
Assuntos
Inseminação Artificial Homóloga , Útero , Muco do Colo Uterino/fisiologia , Coito , Feminino , Humanos , Infertilidade/etiologia , Infertilidade/fisiopatologia , Infertilidade/terapia , Hormônio Luteinizante/sangue , Masculino , Gravidez , Estudos Prospectivos , Espermatozoides/fisiologia , Fatores de TempoRESUMO
Fifty patients (normal responders) received either human menopausal gonadotropin (hMG) alone (control group) or leuprolide + hMG (leuprolide group). The use of leuprolide was associated with a reduction of hMG requirements (14.8 versus 17.8 ampules, P = 0.02) and the abolition of spontaneous luteinizing hormone surges (nil versus 3, P = 0.006). The rate of fertilization (87% versus 65%, P = 0.003) was higher in the leuprolide group. Pituitary and ovarian suppression was effected for 16 subjects who had previously shown a poor follicular response and a further 19 subjects who had previously responded abnormally. The poor responders required more hMG (43.9 versus 27.1 ampules, P less than 0.001), achieved a lower estradiol maximum (5.1 versus 12.1 nmol/l, P less than 0.001), and had fewer oocytes recovered (4.1 versus 11.5, P less than 0.001), than the abnormal responders.
Assuntos
Fertilização in vitro , Transferência Intrafalopiana de Gameta , Hormônio Liberador de Gonadotropina/análogos & derivados , Hormônios/uso terapêutico , Menotropinas/uso terapêutico , Indução da Ovulação , Quimioterapia Combinada , Estradiol/sangue , Feminino , Hormônio Foliculoestimulante/sangue , Hormônio Liberador de Gonadotropina/uso terapêutico , Humanos , Leuprolida , Hormônio Luteinizante/sangue , Oócitos/citologia , Progesterona/sangueRESUMO
The concentrations of the gonadotropins, luteinizing hormone (LH), and follicle-stimulating hormone (FSH) were measured in the luteal phase of the cycle in patients undergoing ovarian hyperstimulation. In nonconception cycles, FSH and LH were increased in the late luteal phase compared with conception cycles in which both gonadotropins were suppressed. Estradiol (E2) and progesterone concentrations increased in pregnancy cycles and may be the sole cause for the decreased gonadotropin concentrations as shown by equivalent concentrations of LH and FSH in both pregnancy and nonpregnancy cycles after matching for E2 concentrations. Subjects who subsequently had twin pregnancy or a spontaneous abortion were compared with those with a successful ongoing singleton conception. There were no significant differences relative to LH and FSH between the three groups, although in twin pregnancy FSH tended to be lower at day 16 from oocyte recovery. It is concluded that suppression of LH and FSH in hyperstimulated pregnancy cycles occurs after the time of the rising human chorionic gonadotropin concentrations in plasma.
Assuntos
Fertilização in vitro , Hormônio Foliculoestimulante/sangue , Transferência Intrafalopiana de Gameta , Hormônio Luteinizante/sangue , Gravidez/sangue , Aborto Espontâneo , Adulto , Estradiol/sangue , Feminino , Humanos , Recém-Nascido , Ciclo Menstrual/sangue , Gravidez Ectópica , Progesterona/sangue , GêmeosRESUMO
The aquaporins (AQ-s) are a group of intrinsic membrane proteins which facilitate movement of water across cell membranes; their recent identification in the kidney has led to the reappraisal of the mechanisms and pathways of water movement across epithelia. Aquaporin-1, (CHIP-28) is reported distributed in cardiac myocytes and vascular smooth muscle cells of large arteries. A related protein, AQ-4, has been identified in the sarcolemma of skeletal muscle fibres. We report aquaporin expression in the cell membrane of smooth muscle cells of the rat genital tract; fluorescence immunohistochemistry of rat uterine (fallopian) tube and vagina demonstrated AQ-1 in visceral smooth muscle of these tissues. In the uterine tube, AQ-1 labelling is most pronounced in the innermost longitudinal and the inner cells of the circular muscle layer and is absent from the outer longitudinal muscle layer of the myosalpinx. The possibility of a specific role for AQ-1 in tubal transport by altering the tubal luminal diameter during the estrus cycle is suggested.
Assuntos
Aquaporinas/análise , Tubas Uterinas/citologia , Músculo Liso/citologia , Vagina/citologia , Animais , Aquaporina 1 , Aquaporina 4 , Membrana Celular/ultraestrutura , Feminino , Imunofluorescência , Técnicas Imunoenzimáticas , Imuno-Histoquímica , Ratos , Ratos WistarRESUMO
Clinical trials were performed between 1987 and 1992 on the use of zona drilling (ZD), zona cutting (ZC) and subzonal sperm microinjection (SZI) for the treatment of severe male infertility. ZD significantly improved the fertilization rate, but embryo morphology was poor and no pregnancies were achieved, so it was abandoned in favour of ZC. The fertilization rate was acceptable in the first trial of ZC but embryo morphology was still poor and no pregnancies were achieved, so a number of protocol changes were instigated. Shrinkage of oocytes in hypertonic sucrose prior to ZC markedly improved embryo quality, whereas transfer of embryos on Day 3 after oocyte retrieval enhanced the pregnancy rate. However, despite these improvements, the overall pregnancy rate in the third ZC trial was still low (16.6% per transfer), so a trial of SZI was initiated in 1992. The overall fertilization rate in 82 SZI cycles was 34.4% and, although the polyspermy rate was high, a clinical pregnancy rate of 30.8% per transfer and an implantation rate of 18.4% per embryo were achieved. These trials demonstrate that SZI is a successful treatment for severe male infertility; under the trial conditions, at least, it was superior to ZD or ZC.
Assuntos
Fertilização in vitro/métodos , Infertilidade Masculina , Microcirurgia/métodos , Interações Espermatozoide-Óvulo , Zona Pelúcida , Desenvolvimento Embrionário e Fetal , Feminino , Fertilização , Humanos , Masculino , Gravidez/estatística & dados numéricosAssuntos
Transferência Embrionária/métodos , Laparoscopia/veterinária , Prenhez , Ovinos/fisiologia , Animais , Feminino , GravidezRESUMO
Oviducal secretions were obtained from conscious unrestrained ewes throughout the oestrous cycle via indwelling cannulae and the content of prostaglandin F (PGF) was determined by radioimmunoassay. Levels of PGF of up to 230 ng/ml were found in oviducal fluids obtained from ewes showing regular patterns of secretion and normal cyclical ovarian function as indicated by plasma progesterone measurement. Relatively large day to day fluctuations in content were evident, but there was no consistent relationship between concentration and stage of the oestrous cycle. Concentrations of PGF in excess of 100 ng/ml were common in preparations where autopsy later revealed infection or tissue irritation, and the concentration of PGF invariably exceeded 75 ng/ml when the concentration of protein in the oviducal fluid was abnormally high.
Assuntos
Líquidos Corporais/análise , Tubas Uterinas/metabolismo , Prostaglandinas F/análise , Ovinos/fisiologia , Animais , Aspirina/farmacologia , Líquidos Corporais/efeitos dos fármacos , Líquidos Corporais/metabolismo , Estro , Feminino , Gravidez , Proteínas/análiseRESUMO
Fewer frozen-thawed mouse oocytes cleaved to the 2-cell stage compared to fresh control oocytes fertilized in vitro (46% vs 79%). The reduced rate of 2-cell formation was only partly explained by a decreased rate of fertilization (63% vs 85%). However, subsequent development to expanded blastocysts was not different (75% vs 78%). An increased frequency of second polar body retention by fertilized frozen-thawed oocytes compared with controls (11.8% vs 1.3%) was shown to be largely responsible for the higher incidence of polyploidy (16.3% vs 3.7%). The frequency of polyspermic fertilization was not different in the two groups (3.9% vs 2.3%).
Assuntos
Fertilização in vitro , Oócitos/ultraestrutura , Poliploidia , Preservação de Tecido , Animais , Blastocisto , Células Cultivadas , Fase de Clivagem do Zigoto , Congelamento , CamundongosRESUMO
The qualitative profiles of the proteins synthesized by sheep oocytes at various stages of maturation were determined by electrophoretic separation in one dimension on polyacrylamide SDS gels. No change in protein synthetic pattern was observed in ooce changes had taken place in at least 12 separate protein bands. Marked alterations in the synthesis of some proteins were apparent 15 h after LH; formation of proteins in 5 of the original bands was either reduced or not detectable, while new synthesis was evident from the appearance of 7 additional bands. The pattern of proteins produced by oocytes cultured within the follicle corresponded closely with that observed in vivo: changes in synthesis were initiated about 9 h after addition of gonadotrophin and were completed by 15 h. Oocytes cultured outside the follicle in a gonadotrophin-containing medium did not exhibit a change in protein synthesis and at 15 h only those proteins produced during the early stages of maturation were being synthesized.
Assuntos
Oócitos/metabolismo , Óvulo/metabolismo , Biossíntese de Proteínas , Animais , Células Cultivadas , Eletroforese em Gel de Poliacrilamida , Feminino , Hormônio Liberador de Gonadotropina/farmacologia , OvinosRESUMO
Levels of reproductive steroids and gonadotrophins were analysed retrospectively during the peri-implantation period following non-conceptional and conceptional natural cycles and in cycles associated with ovarian hyperstimulation for in-vitro fertilization or gamete intra-Fallopian transfer. In cycles not associated with conception, the luteal phase of hyperstimulated cycles (n = 100) was characterized by higher serum progesterone and oestradiol levels (P less than 0.01) and with an earlier decline in steroids than in natural cycles (n = 21). On day 11 (day of oocyte recovery = day 0), the level of progesterone in twin (n = 59) and triplet (n = 13) pregnancies was higher than singleton pregnancies (n = 176) (P less than 0.006, P less than 0.006 respectively) while those destined to abort (n = 66) had lower progesterone levels (P less than 0.01). Ectopic implantation (n = 11) had the lowest progesterone concentrations on day 11 (P less than 0.01) and this may imply a delay in corpus luteum rescue or a later implantation time than intrauterine conception.
Assuntos
Corpo Lúteo/fisiologia , Estradiol/sangue , Gravidez Ectópica/sangue , Gravidez Múltipla/sangue , Progesterona/sangue , Clomifeno/farmacologia , Feminino , Fertilização in vitro , Transferência Intrafalopiana de Gameta , Humanos , Fase Luteal/fisiologia , Hormônio Luteinizante/sangue , Menotropinas/farmacologia , Ovário/efeitos dos fármacos , Gravidez , Resultado da Gravidez , Estudos RetrospectivosRESUMO
When there is a risk of inherited disease, preimplantation diagnosis gives couples an opportunity to avoid having a child with the disease. Sex determination can be used to exclude the likelihood of a sex-linked disorder. Accuracy of the diagnosis is important. We have tested the reliability of sex determination based on the recognition of a testis-determining gene (SRY) sequence. DNA from the blood of 120 men and women and from 38 single lymphocytes was amplified by polymerase chain reaction (PCR) with the SRY and control (ZP3) gene primers. All results confirmed the correct sex of origin (100%). The test was then used to determine the sex of 21 single embryo cells biopsied from 21 (4-8 cell) human polyspermic embryos. 2 embryo cells recognised at biopsy to have degenerated produced negative results. The other 19 single embryo cells showed 100% PCR amplification. 11 (58%) of the embryos were judged to be "male" and 8 (42%) "female". The SRY and ZP3 gene primers selected are highly specific and give accurate results in sex determination and their use provides a new reliable method for routine preimplantation and general prenatal sex determination in man.
Assuntos
Blastocisto , DNA/genética , Reação em Cadeia da Polimerase , Análise para Determinação do Sexo/métodos , Cromossomo Y , Animais , Sequência de Bases , Blastômeros , Primers do DNA , Feminino , Humanos , Linfócitos , Masculino , Dados de Sequência Molecular , GravidezRESUMO
In this report, we present the results of our first 100 consecutive cycles of intracytoplasmic sperm injection (ICSI). Overall, fertilization occurred in 98% of cycles and embryos were transferred in 94% (2.6 embryos per cycle). About 50% of patients had embryos frozen. The overall fertilization rate was 71%, of which 4% were abnormally fertilized (three pronuclei). A total of 30 clinical pregnancies were established (32% per transfer), resulting in 18 singleton, six twin and one triplet ongoing pregnancies. The implantation rate per embryo was 15%. There were no significant differences in the fertilization or pregnancy rates between patients who had only occasional motile spermatozoa in the ejaculate, semen that was too poor for routine in-vitro fertilization (IVF), or who had failed routine IVF and/or subzonal sperm injection (SUZI). A group of 18 patients were treated with both ICSI and routine IVF on their first cycle because of the high likelihood of failed fertilization due to poor sperm morphology < 20% normal). In this group, ICSI oocytes had a fertilization rate of 76% compared to only 15% for the routine IVF (control) oocytes, and six patients conceived after transfer of ICSI embryos (33%), indicating that ICSI can be used successfully on 50% of the oocytes if fertilization failure is expected. Similarly, patients who had failed to become pregnant with SUZI achieved excellent results after ICSI. There were no significant differences between ICSI and routine IVF in the proportions of grade 1, 2 or 3 embryos on day 3 post-oocyte recovery.(ABSTRACT TRUNCATED AT 250 WORDS)