Microanatomy of blood vessels in spermatic cords and its clinical implication / 中华男科学杂志

<p><b>OBJECTIVE</b>Both microsurgical subinguinal varicocelectomy (MSIV) and microsurgical high inguinal varicocelectomy (MHIV) are recommended for the treatment of varicocele, but they differ in technical complexity. This study aimed to determine the microanatomy of spermatic blood vessels in the two surgical approaches.</p><p><b>METHODS</b>We recorded the numbers of spermatic veins, arteries and lymphatics in 80 cases of MSIV and 20 cases of MHIV. We also examined the spermatic cords from 10 adult male cadavers by histological staining.</p><p><b>RESULTS</b>The numbers of medium spermatic veins (2 -5 mm in diameter) were 1.80 +/- 0.83 and 3.98 +/- 1. 99 in MHIV and MSIV, respectively, with significant difference between the two groups (t = -7.536, P < 0.01), and the total numbers of spermatic veins were 6.40 +/- 1.67 and 9.01 +/- 2.70, also with significant difference between the two (t = -4.071, P < 0.01). However, there were no significant differences between MHIV and MSIV in the numbers of small spermatic veins (diameter < or = 2 mm), large spermatic veins (diameter > or = 5 mm), arteries and lymphatics, nor in the numbers of spermatic veins and arteries of the cadavers.</p><p><b>CONCLUSION</b>The total number of spermatic veins and the number of medium spermatic veins may be larger in MSIV than in MHIV, but the medium spermatic veins do not increase surgical difficulty, and MSIV is not more complicated than MHIV.</p>

[ retrospective study of 339 ICSI cycles: assessment of the two first years of activity of the assisted reproductive technology's center of Aziza Othmana's hospital]

Intracytoplasmic sperm injection [ICSI] is a micro-manipulation-assisted fertilization, whereby one spermatozoon is injected into the oocyte cytoplasm. Initially, ICSI was the treatment of choice for male factor infertility. However, because of the high fertilization and pregnancy rates achieved with this technique, the scope of the procedure has been widened to include couples with other causes of infertility. The aim of this study was to study the progression of the activity of the assisted reproductive technology's center of Aziza Othmana's Hospital and the ICSI results during the first two years. Our study included 269 infertile couples who underwent 339 ICSI cycles between 1st May 2001 and 30 April 2003. Cycles with no oocytes obtained at the follicular aspiration and women aged over 40 years were excluded from this study. The number of ICSI cycles progressed in our center: 150 ICSI cycles in the 1st year, 189 ICSI cycles in the 2nd year. The mean number of picked-up oocytes was 8,8 +/- 5,6. The fertilization rate was 62. The mean number of transferred embryos was 3,1 +/- 1,5. The pregnancy rate per transfer was 32,4%. The miscarriage rate was 28,4%. The take home baby rate was 67,9%. The number of couples undergoing ICSI cycles in our center is increasing. The fertilization rates and pregnancy rates in our center are similar to those published in the literature

In Vitro Fertilization Program

Rapid progress has been made in the field of infertility since the first IVF (in vitro fertilization) baby was born in 1978. Controlled ovarian stimulation with FSH is currently the standard procedure for ovarian stimulation before follicular aspiration. Gonadotropin-releasing hormone agonists and antagonists have been used to prevent endogenous LH surge during controlled ovarian hyperstimulation.The goal of controlled ovarian stimulation with gonadotropins is to obtain a large number of mature oocytes and thereby improve the likelihood of obtaining an adequate number of embryos for subsequent transfer. IVF was initially presented as a treatment for tubal factor infertility but was quickly utilized in other areas in the field of infertility, such as male factor infertility and even ovarian failure. ICSI (intracytoplasmic sperm injection) is a more recent approach for male factor treatment, which allows the sperm to be directly injected into the egg using micromanipulation. Preimplantation genetic diagnosis can be performed on embryos prior to the embryo transfer. The complications associated with the IVF program include ovarian hyperstimulation syndrome and multiple pregnancies. The multiple pregnancies are directly related to the practice of transferring multiple embryos at embryo transfer. Each IVF clinic publishes its pregnancy rates. However, comparisons between clinics are difficult because the success rates vary depending on the distribution of underlying causes and age of the patients. The current take-home-baby rate is only 34.7%. In 2005, the Korean government enacted a law to regulate many aspects of IVF practice.

Biochip micromachining technology adopting laser technology / 生物医学工程学杂志

This paper deals with the manufacturing state of the art of biochip, and introduces a new method--laser microtechnology, including its developing procedure, characteristics and function in biochip production.

Diseño y construcción de un prototipo de micromanipulador / Design and construction of a micromanipulator prototype

Este proyecto se hizo con el fin de proveer a la comunidad científica de un micromanipulador de alta calidad, de bajo costo y útil para manipular células con fines de investigación y aplicación biológica. Los procedimientos que se llevaron acabo fueron: - Diseño del prototipo del micromanipulador, de fácil manejo para ser utilizado en técnicas de reproducción asistida.- Estudio de los posibles materiales, su disponibilidad, su durabilidad y su resistencia.- Análisis del costo de los materiales, del diseño, de la mano de obra y su relación costo beneficio.- Construcción del micromanipulador y realización de diferentes pruebas de su utilidad y aplicación en animales, u otros experimentos biológicos no humanos. Se logro obtener una herramienta para técnicas de reproducción asistida al que, además, puedan integrársele posteriormente nuevos dispositivos para las múltiples aplicaciones...

The analysis of fetal gender and BclI polymorphism with fetal cells in maternal blood / 대한산부인과학회잡지

OBJECTIVE: We used nucleated erythrocytes in maternal blood for prenatal determination of the fetal gender as the preliminary experiment for the screening of fetal genetic status and the BclI DNA polymorphism in an attempt to clarify the origin of erythrocytes in maternal blood. METHODS: In seventeen pregnant women, venous blood was withdrawn and the nucleated erythrocytes were recovered by magnetic activated cell sorting (MACS) and immunostaining. After isolation of nucleated erythrocytes by micromanipulation, we performed nested PCR for amelogenin gene to identify the fetal gender and performed BclI DNA polymorphism to clarify the origin of erythrocytes. RESULTS: We could amplify the minute DNA in a single cell by primer extension preamplification and nested PCR of amelogenin gene in 94 (48.7%) cells and could identify the fetal gender by 58.8%. BclI DNA polymorphism revealed that the several cells, which did not reveal the specific band of Y chromosome in spite of the pregnancy of male fetuses, must be the cells from mother. CONCLUSION: Through this study, we could conclude that several nucleated erythrocytes in maternal blood circulation can originate from mother, therefore we must develop the new method to identify the nucleated erythrocyte of fetal origin. Considering that we must apply for the larger number of pregnant women to screen, the procedure was multi-step and complex. Therefore, we must design the new scheme to utilize the nucleated erythrocytes in maternal blood.

Developmental competence and Effects of Coculture after Crypreservation of Blastomere-Biopsied Mouse Embryos as a Preclinical Model for Preimplantation Genetic Diagnosis / 대한불임학회지

OBJECTIVE: The effects of cryopreservation with or without coculture on the in vitro development of blastomere-biopsied 8-cell mouse embryos were investigated. This experimental study was originally designed for the setup of a preclinical mouse model for the preimplantation genetic diagnosis (PGD) in human. METHODS: Eight-cell embryos were obtained after in vitro fertilization (IVF) from F1 hybrid mice (C57BLfemale symbol/CBAmale symbol). Using micromanipulation, one to four blastomeres were aspirated through a hole made in the zona pellucida by zona drilling (ZD) with acid Tyrode's solution (ATS). A slow-freezing and rapid-thawing protocol with 1.5M dimethyl sulfoxide (DMSO) and 0.1M sucrose as cryoprotectant was used for the cryopreservation of blastomere- biopsied 8-cell mouse embryos. After thawing, embryos were cultured for 110 hours in Ham's F-10 supplemented with 0.4% bovine serum albumin (BSA). In the coculture group, embryos were cultured for 110 hours on the monolayer of Vero cells in the same medium. The blastocyst formation was recorded, and the embryos developed beyond blastocyst stage were stained with 10% Giemsa to count the total number of nuclei in each embryo. RESULTS: The survival rate of embryos after cryopreservation was significantly lower in the blastomere-biopsied (7/8, 6/8, 5/8, and 4/8 embryos) groups than in the non-biopsied, zona intact (ZI) group. Without the coculture, the blastocyst formation rate of embryos after cryopreservation was not significantly different among ZI, the zona drilling only (ZD), and the balstomere-biopsied groups, but it was significantly lower than in the non-cryopreserved control group. The mean number of cells in embryos beyond blastocyst stage was significantly higher in the control group (50.2+/-14.0,) than in 6/8(26.5+/-6.2), 5/8(25.0+/-5.5), and 4/8(17.8+/-7.8) groups. With the coculture using Vero cells, the blastocyst formation rate of embryos after cryopreservation was significantly lower in 5/8 and 4/8 groups, compared with the control, 7/8, and 6/8 groups. The mean number of cells in embryos beyond blastocyst stage was also significantly lower in 4/8 group (25.9+/-10.2), compared with the control (50.2+/-14.0), 7/8 (56.0+/-22.2), and 6/8 (55.3 +/-25.5) groups. CONCLUSION: After cryopreservation, blastomere-biopsied mouse embryos have a significantly impaired developmental competence in vitro, but this detrimental effect might be prevented by the coculture with Vero cells in 8-cell mouse embryos biopsied one or two blastomeres. Biopsy of mouse embryos after ZD with ATS is a safe and highly efficient preclinical model for PGD of human embryos.

Análise crítica da injeçäo intracitoplasmática de espermatozóides (ICSI) / Critical analysis of intracytoplasmatic sperm injection (ICSI)

Recentes estudos propiciaram um melhor entendimento da fisiologia reprodutiva, com conseqüente evoluçäo das técnicas de reproduçäo assistida. Após a fertilizaçäo in vitro (FIV) surgiram outras técnicas de fertilizaçäo assistida tais como a injeçäo intracitoplasmática de espermatozóides (ICSI), introduzida em 1992, que consiste na injeçäo de um único espermatozóide no citoplasma de um oócito com o auxílio de uma micropipeta. Esta nova técnica aumentou dramaticamente as taxas de sucesso nos casos de infertilidade masculina e tornou-se em todo mundo a técnica de micromanipulaçäo de escolha. De acordo com a literatura, näo há diferenças nas taxas de fertilizaçäo e gravidez entre a ICSI e a FIV quando esta é realizada na ausência do fator masculino de infertilidade. Entretanto, na presença de qualquer fator masculino de infertilidade, a ICSI tem se mostrado nitidamente superior. O desenvolvimento da ICSI levantou inúmeras questöes no que se refere aos riscos genéticos envolvidos. Evidências recentes demonstram que a evoluçäo das gestaçöes e a taxa de malformaçöes congênitas pós-ICSI parecem ser semelhantes às obtidas com outras técnicas de reproduçäo assistida. Somente o seguimento cuidadoso e contínuo das gravidezes e dos neonatos poderá responder definitivamente esta questäo.

Fertilizaçäo assistida em más-respondedoras: até quando prosseguir? / Assisted reproduction in poor-responders: how far to go?

OBJETIVOS: analisar a influência de um acréscimo substancial na administraçäo de gonadotropinas durante a induçäo ovulatória em ciclos FIV clássica/ICSI sobre os resultados finais. CASUISTICA: pacientes em tratamento de infertilidade que se submeteram a ciclos de FIV clássica/ICSI de jan/94 a dez/95 no Serviço de Infertilidade do Hospital Antoine Béclère - Clamart/Paris. PLANO DE ESTUDO: Análise retrospectiva. MÉTODOS: Foram analisados 484 ciclos no total, 325 FIV clássica e 159 ICSI. Destes ciclos, 62 tinham níveis de estradiol no 6§ dia de induçäo abaixo de 60 pg/ml. Estes ciclos foram divididos em ciclos de grande requisiçäo hormonal final para se obter uma resposta adequada ou näo (corte em 60 ampolas finais totais de gonadotropinas). As taxas de gravidez nestes grupos foram analisadas. RESULTADOS: Os ciclos nos quais menos de 60 ampolas totais de gonadotropinas foram empregadas e que menos de 6 ampolas eram administradas no 6§ dia obtiveram melhor performance, com maior número de gravidezes estatisticamente significante.

Micromanipulation and cloning studies on buffalo oocytes and embryos using nucleus transfer.

An investigation for testing the viability of production of cloned buffalo embryos through nucleus transfer has been made. Matured buffalo oocytes, after zona cutting to an extent of 60 degrees near polar body, were enucleated using a new approach. Instead of aspirating the cytoplasm contents in a pipette, the half of cytoplasm of oocyte was pushed out, thereby also taking away the nuclear material of the oocyte, leaving the demi-oocyte with the zona pellucida enucleated. The absence of fluorescence confirmed the success of the enucleating process. For enucleating, the oocytes which had intact plasma membrane were eligible for bisectioning. There was no significant difference in oocytes having intact membrane among grade I (33.9%) and grade II (31.4%) oocytes, whereas lower percentage of grade III oocytes had a very low percentage having intact plasma membranes (8.5%). The hours of maturation for 32, 37 and 42 did not influence the per cent oocytes which had intact membranes. All the bisected or demi-oocytes tested with fluorescence screening yielded successful enucleation in 88.2% demi-oocytes. The temporal effect of three maturation hours of 32, 37, and 42 hr; two electrical pulse numbers of 2 and 3 pulses and two magnitudes of electric pulses of 15 and 20 V were studied for their effect on the percentage of successful fusion of demi-oocyte blastomere complexes and the rate of complexes undergoing cleavages. The time period for which the oocytes were subjected to the process of maturation significantly affected the per cent fusions and per cent cleavage of the demi-oocyte blastomere complexes and 32 hr maturation yielded less fusions (38.5%) compared to maturation for 37 and 42 h (53.2 and 57.8%, respectively). The treatment of either 2 or 3 electrical pulse numbers resulted in significantly different fusion (45.6 and 54.1%) as well as cleavage rates (18.2 and 26.1%) of demi-oocyte-blastomere complexes electrofused. The treatment of two levels of magnitude of 15 and 20 V of an electric current resulted in similar per cent fusion (48.0 and 51.6%) and cleavage rates (21.0 and 23.2%). Fortified TCM with either 10 or 20% FBS for culturing freshly electrofused complexes for 1 hr did not differ significantly with respect to per cent complexes fused and cleaved, giving a fusion rate of 46.2 and 53.8% and cleavage rate of 21.2 and 23.2% for 10% and 20% FBS, respectively. Production of cloned embryos through the process of nuclear transfer has been accomplished. The successful cleavages of the nuclear transferred oocytes demonstrated the viability of enucleation procedures of the oocytes and technology implementation of electrofusion in buffalo oocytes.

Microassisted Fertilization of Human Oocytes with Intracytoplasmic Sperm Injection in In VitroFertilization Program / 대한산부인과학회잡지

In spite of much progress in in vitro fertilization and embryo transfer(IVF-ET) program,the pregnancy rate remains at 20~30%, and the endometrial implantation rate per embryotransferred at 10%. Although IVF-ET is widely applied in the treatment of coupleswith male factor infertility, it may fail in many infertile couples with normal semen parameters,and certain couples cannot be accepted for standard IVF-ET due to unfertilization orextremely low fertilization rate of oocytes. Recently, several procedures of microassistedfertilization(MAF) using micromanipulation have been introduced, and pregnancies and birthshave been obtained after intracytoplasmic sperm injection(ICSI).This clinical study was performed to develop and establish ICSI as an effective procedureof MAF in infertile couples who could not be accepted for standard IVF-ET becauseof extremely impaired semen characteristics(Group A) and because of failure in fertilizationof extremely low fertilization rate of oocytes with the conventional fertilization technique inthe previous IVF-ET cycles(Group B). From March, 1995 to December, 1996, a total of 114cycles of IVF-ET with ICSI in 65 infertile patients were included in study group, and theoutcomes of ICSI were analyzed according to fertilization rate, cumulative embryo score(CES), and pregnancy rate.In Group A, 34 patients were evaluated with semen score such as number of totalmotile sperms, and then divided into 4 groups accordingly. In 62 ICSI cycles, the numberof oocytes retrieved after controlled ovarian hyperstimulation(COH) was 12.4+/-6.8, and thenumber of oocytes optimal for ICSI procedure was 8.8+/-5.5. The fertilization rate of 65.7+/-23.6% could be obtained after ICSI. The number of embryos transferred was 4.4+/-2.2 withthe mean CES of 50.5+/-34.3 in ICSI cycles. The overall pregnancy rate was 24.2%(15/62)per cycle and 44.1%(15/34) per patient. There were no significant differences in the pregnancyrates among 4 groups. Although more mature oocytes were retrieved, the fertilizationrate was significantly lower in Group A-1 compared with Group A-IV. However, semenscore did not clearly affect the outcomes of ICSI in couples with severe male factor infertility.In Group B, the number of oocytes retrieved after COH was 10.5+/-6.1 in 49 previouscycles, and 10.8+/-5.7 in 52 ICSI cycles. In ICSI cycles, the number of oocytes optimal forICSI procedure was 8.5+/-5.1 with the fertilization rate of 72.4+/-22.5%. The number ofembryos transferred was 1.4+/-2.4 in previous cycles, and 4.7+/-1.8 with the mean CES of 50.4+/-29.9 in ICSI cycles. In ICSI cycles, the overall pregnancy rate was 30.8%(16/52) percycles and 51.6%(16/31) per patients.In conclusion, MAF of human oocytes with ICSI is a promising fertilization method forIVF-ET patients, especially with few spermatozoa for the conventional methods of in vitroinsemination and with the past history of failure in fertilization or low fertilization rate inthe previous cycles, and ICSI using micromanipulation procedures applied to human oocyteswill provide a range of novel techniques which may dramatically improve the pregnancy ratein IVF-ET program and contribute much to the effective management of infertile couples.

Impreovement in Endometrial Implantation of Humman Embryos with Assisted Hatching Using Micromanipulation in Intrauterine Embryo Transfer / 대한산부인과학회잡지

Although the fertilization rate exceeds to 80~90% with much progress in vitro fertilizaton and embyo transfer(IVF-ET) program, the prgnancy rate rmains at 20~30%, and the endometrial implantaion rate per embryo transferred at 10~15%. Recently, many attempts have been made to improve embrynic implantion after IVF-ET including serveral procedures of assisted hatching(AH) using micromanipulation, and pregnacies and births have been obtained after AH. This clinical study was performed to develop and estabilish AH as an effective procedure to improve embryonic implantioan in IVF-ET patients who had previous repeated failure of standard IVF-ET more than 2 times(Group R), were more than 37 years old(Group A), or had high basal serun FSH levels more than 15 mIU/ml(Group F). From January, 1995 to Februry, 1996, 132 cycles of AH using partial zona dissection(PZD) were performed in 104 infertile patients, and the outcomes of AH were analyzed according to pregnancy rate. The number of oocytes retrieved after controlled ovarian hyperstimulation(COH) was 9.9+/-7.1 in 71 cycles of 54 patients who had previous repeated failure more than two times(Group I: Group R,R+A,R+F, and R+A+F), 8.4+/-5.9 in 62 cycles of 46 patients whose age was more than 37 years old(Group II : Groups A, R+A, A+F, and R+A+F), and 8.7+/-6.5 in 49 cycles of 47 patients who had high basal serum FSH levels more than 15 mIU/ml(Group III:Groups F,R+F, A+F, and R+A+F). The number of embroys transferred after AH was 4.7 +/-1.8 in Group I, 4.2 +/-1.9 in Group II, and 4.2+/-2.0 in Froup III. The mean cumulative embryo score(CES) was 56.8+/-30.0 in Group I, 52.6+/-30.6 in Group II, and 52.6+/-29.9 in Group III. There were no significant differences in the numbers of oocytes rerieved and embryos transferred, and CES among 3 groups. The overall clinical pregnancy rate was 14.4%(19/132) per cycle and 18.3%(19/104) per patient. THe clinical pregnancy rate per cycle and per patient was 12.7%(9/71) and 16.7%(9/54) in Group I, 4.8% (3/62) and 6.5%(3/46) in Group II, and 26.5%(13/49) and 27.7%(13/47) in Group III, and there was a significant difference between Group II and Group III. In conclusion, AH of human embryos using micromanipulation might be promising for IVF-ET patients, especially with the past history of repeated failure, old age, and high basal serum FSH level and AH will provide a range of novel techiques which may dramatically improve the implanatation and pregnancy rates in IVF-ET program and contribute much to effective management of infertile couples.

Injeçäo intracitoplasmática de espermatozóides em oócitos humanos näo fertilizados, 1 dia após inseminaçäo in vitro / Intracytoplasmic sperm injection for re-insemination of oocytes that failed fo fertilize after in vitro fertilization

A reduçäo nas taxas de fertilizaçäo, determina um menor número de embriöes disponíveis para transferência. Para evitar este tipo de problema, a re-inseminaçäo dos oócitos näo fertilizados tem sido sugerida, pela técnica de ICSI. O objetivo do trabalho foi identificar a capacidade de fertilizaçäo de oócitos näo fertilizados 1 dia após FIV convencional, pela técnica de ICSI, e observar o potencial de gravidez dos embriöes formados. Um total de 67 oócitos foram aspirados das 5 pacientes com infertilidade primária incluídas no estudo. Deste total, 46 (68,5 por cento) foram selecionados para ICSI um dia após a inseminaçäo. Dos 46 oócitos submetidos a ICSI, 26 apresentaram 2 pró núcleos (56 por cento) e 21 apresentaram divisäo, com uma taxa de clivagem de 80,7 por cento. Das 4 pacientes que obtiveram embriöes transferidos no dia 3 pós-punçäo folicular, 2 apresentaram níveis de bHCG sérico igual a 25 UI/ml, 12 dias após a TE. Os resultados säo prematuros para se afirmar a confiabilidade do método, mas a taxa de fertilizaçäo, a qualidade dos embriöes obtidos, e a presença de duas gestaçöes nos 5 ciclos realizados, parece nos mostrar uma opçäo para os casos de ausência completa de fertilizaçäo em ciclos de FIV.

Punçäo testicular com agulha fina seguida de injeçäo intracitoplasmática de espermatozóide e gravidez: relato de caso / Fine needle biopsy of the testis followed by intracytoplasmic injection of the sperm and pregnancy: case report

Nos últimos anos, com o grande avanço das técnicas de reproduçao assistida, já se tornou possível a obtençao de gravidez utilizando-se apenas um espermatozóide, o qual pode ser injetado diretamente no interior do citoplasma do oócito (ICSI). Recentemente, a obtençao de espermatozóides através de punçao testicular com agulha fina nos mostrou que estes, apesar de imóveis e imaturos, sao capazes de fertilizar e resultar em gestaçao viável. Com isto, os pacientes portadores de fator masculino severo adquiriram uma nova esperança terapêutica. Apresentamos a seguir um caso de gravidez viável obtida com espermatozóide aspirado com agulha fina diretamente do testículo e ICSI.

Embarazo con espermatozoides testiculares / Pregnacy with testicular spermatozoa

En un paciente con diagnóstico de agenesia de vías espermáticas se propuso afectuar una aspiración epididimaria asociada a FIV (fertilización in vitro). En la exploración excrotal no se encontró epidídimo (debido probablemente a antecedente quirúrgico de biopsia epididimaria) por lo que se efectuó una biopsia testicular. Este material fue procesado obteniéndose escasos espermatozoides (algunos cientos). Estos espermatozoides fueron inyectados en el citoplasma de los ovocitos de la esposa (técnica de ICSI).De los 18 ovocitos inyectados, 3 se dañaron durante el procedimiento y a las 16 horas se observaron signos de fertilización normal en 6 de ellos, prosiguiendo el desarrollo 4 de embriones que fueron transferidos al útero por vía transcervical, 3 en estado de 8 células y uno de 6 células, habiéndosele efectuado previamnete en la zona pelúcida de cada uno de los embriones un orificio de 20 micrones con solución ácida de Tyrode. A los 15 días de la transferencia (2-1-95) el resultado de la subunidad beta de HCG fue positiva, detectándose al mes (2-2-95) por ecografía un embrión de 19 mm, cursando un embarazo normal

Gravidezes com micromanipulaçäo a partir de ovocitos e pré-embriöes transportados para laboratório central especializado / Pregnancies obtained after micromanipulation of oocytes and embryos transported to a specialized center

OBJETIVOS: Avaliar os resultados obtidos com a injeçao intracitoplasmática de espermatozóides em ovocitos que tenham sido submetidos a transporte de centro de fertilizaçao em vitro clássica, para outro especializado em micromanipulaçao. MATERIAL E MÉTODOS: 18 pacientes foram submetidas a superovulaçao com GnRH(a), FSH e HCG. Foram obtidos 145 ovocitos,os quais foram transportados em câmara especial, para centro especializado em micromanipulaçao. O sêmen foi preparado pela técnica de gradiente descontínuo de Percoll. Os ovocitos foram denudados com 80U/ml de hialuronidase. A injeçao intracitoplasmatica foi realizada em microgotas sob óleo mineral, utilizando equipamento Narishige. RESULTADOS: Após a injeçao intracitoplasmática, 35 dos 145 ovocitos fertilizaram (2 pró-nucleos), 37 sofreram lise, 73 evoluíram de forma atípica e houve 5 gestaçoes clínicas. CONCLUSOES: A micromanipulaçao é restrita a alguns centros devido às dificuldades técnicas e custo elevado do equipamento. Sendo o transporte fator que nao interfere de maneira significativa nos resultados deste procedimento, sugerimos a criaçao de serviços centralizados, especializados em micromanipulaçao, fazendo com que pelo menos os pacientes com fator masculino grave se beneficiem com estas técnicas.

Embarazo con espermatozoides testiculares / Pregnancy with testicular spermatozoa

En un paciente con diagnóstico de agenesia de vías espermáticas se propusó efectuar una aspiración epididimaria asociada a FIV (fertilización in vitro). En la exploración escrotal no se encontró epidídimo (debido probablemente a antecedente quirúrgico de biopsia epididimaria) por lo que se efectuó una biopsia testicular. Este material fue procesado obteniéndose escasos espermatozoides (algunos cientos). Estos espermatozoides fueron inyectados en el citoplasma de los ovocitos de la esposa (técnica de ICSI). De 18 ovocitos inyectados, 3 se dañaron durante el procedimiento y a las 16 horas se observaron signos de fertilización normal en 6 de ellos, prosiguiendo el desarrollo 4 embriones que fueron transferidos al útero por vía transcervical, 3 en estado de 8 células y uno de 6 células, habiendose efectuado previamente en la zona pelúcia de cada uno de los embriones un orificio de 20 micrones con solución ácido de Tyrode. A los 15 días de la transferencia (2-1-95) el resultado de la subunidad beta de HCG fue positiva, detectándose al mes (2-2-95) por ecografía un embrión de 19 mm, cursando un embarazo normal