Intracytoplasmic Sperm Injection Program Research Articles

Development and Evaluation of Single Sperm Washing for Risk Reduction in Artificial Reproductive Technology (ART) for Extreme Oligospermic HIV Positive Patients

The serodiscordant couples, where the male is HIV-positive, are treated in fertility clinics, using the sperm washing technique by gradient centrifugation. This protocol cannot be carried out in oligo-azoospermic patients, where spermatozoa retrieval from the epididymis and testis must be performed. We developed a single sperm washing technique, where the spermatozoa, after the retrieval, are washed with the aid of a micromanipulator, to obtain virus decontamination and then used for the intracytoplasmic sperm injection (ICSI). The experiment was performed by using sperm samples containing three different viral loads. After one hour of incubation, spermatozoa were taken one by one from the HIV loaded drop and washed in four different microdrops. Before each passage into the next washing drop, the pipette was emptied in a first waste drop and then loaded with new washing medium from a second separate loading drop. After transferring of 10 spermatozoa in these four successive drops, the washing medium and the virus-loaded drops were tested for the HIV RNA presence by the nested RT-PCR technique. The presence of the virus was detected in the waste drop of all three viral loads. The four washing microdrops were each time negative for the presence of HIV-1 RNA, tested by the nested RT-PCR technique. The results show that by rinsing the spermatozoa four times, we are able to diminish the viral load to an undetectable level. Our data demonstrate that single sperm washing can be performed in the cases of extreme male sterility in HIV-positive men. From now on the couples, where the male is oligoazoospermic and HIV positive, could be included in our ICSI program, respecting the usual viral safety level of the ART techniques for the embryo.

Obesity and increased body fat composition significantly affect intracytoplasmic sperm injection (ICSI) outcomes

OBJECTIVE: Obesity is a rising health problem worldwide that is known to impair human reproduction and considered as a contributing factor for infertility. its impact on all outcomes of ICSI program among infertile women was assessed in this study.

Involvement of macrophage colony-stimulating factor (M-CSF) in the function of follicular granulosa cells

To evaluate the role of macrophage colony-stimulating factor (M-CSF) in steroidogenesis by luteinized granulosa cells (GCs) and to examine the effect of FSH, E(2), and P on the production of M-CSF by GCs. In vitro experimental study. University hospital with an IVF-intracytoplasmic sperm injection program. Forty infertile women who underwent intracytoplasmatic sperm injection. The GCs obtained from these patients during oocyte retrieval were cultured and then treated with different concentrations of M-CSF, FSH, E(2), or P for 72 hours. The M-CSF concentrations were determined by ELISA, and the concentrations of E(2) and P were measured by enzyme immunoassay. The M-CSF secretion of GCs was enhanced by FSH or E(2) in vitro in a dose-dependent manner but was unaffected by P. On the other hand, M-CSF elicited a production of E(2) and P by GCs in a dose-dependent manner in the presence or absence of FSH. The results suggest that the growth factor M-CSF might have an important role in ovarian function.

Seminal Plasma Cobalamin Significantly Correlates With Sperm Concentration in Men Undergoing IVF or ICSI Procedures

Mild hyperhomocysteinemia is caused by B vitamin deficiencies. We hypothesize that these biochemical derangements detrimentally affect spermatogenesis. Therefore, the aim of this study was to investigate the folate, cobalamin, pyridoxine, and homocysteine concentrations in blood and seminal plasma and the associations between these biomarkers and semen parameters in men participating in an in vitro fertilization or intracytoplasmic sperm injection program. From 73 men (median age [range]: 37 years [28-53]), blood and semen samples were obtained for the determination of serum and red blood cell (RBC) folate, serum total cobalamin, whole-blood pyridoxal-5'-phosphate, plasma total homocysteine (tHcy), and serum total testosterone. Semen analysis included sperm concentration, motility, and morphology according to World Health Organization criteria. The B vitamins and tHcy concentrations were significantly correlated in blood but not in seminal plasma. The serum and RBC folate concentrations were significantly correlated also with the total folate concentration in seminal plasma (r = .44; P < .001 and r = .39; P < .001, respectively). Likewise, the total cobalamin concentration in serum and seminal plasma was significantly correlated (r = .55; P = .001). Of interest is that the total cobalamin concentration in seminal plasma was significantly correlated with the sperm concentration (r = .42; P < .001). This is in contrast to the absence of significant associations between the other vitamins and tHcy in blood and seminal plasma and any of the semen parameters. These findings suggest that folate and cobalamin are transferred from the blood to the male reproductive organs and emphasize the role of cobalamin in spermatogenesis in human.

Human albumin does not prevent ovarian hyperstimulation syndrome in assisted reproductive technology program: a prospective randomized placebo-controlled double blind study

We aimed to clarify the efficiency of IV human albumin in the prevention of ovarian hyperstimulation syndrome (OHSS). We found that human albumin at the described strength does not seem to either prevent or reduce the incidence of severe OHSS in high risk patients undergoing intracytoplasmic sperm injection (ICSI).

Evaluation of reproductive potential after intracytoplasmic sperm injection of varied human semen tested by antiacrosomal antibodies

To determine whether varied human spermatozoa, as detected with monoclonal antibodies against acrosomal proteins, have an influence on fertilization, transfer, pregnancy, and implantation rates when intracytoplasmic sperm injection is used. A retrospective study. A private IVF center and academic research laboratory. One thousand two hundred forty men participating in the intracytoplasmic sperm injection program. Sperm were divided into seven groups: oligozoospermia, oligoasthenozoospermia, and oligoasthenoteratozoospermia and fresh and frozen-thawed epididymal and fresh and frozen-thawed testicular sperm. Fertilization, transfer, pregnancy, and implantation rates were recorded in each category. Sperm were tested with antibodies for detection of the of the sperm acrosome. Fertilization, transfer, pregnancy and implantation rates, and percentage of acrosome-reacted cells. The fertilization rate and statistical evaluation showed differences between morphologically normal and pathological sperm and other groups. The freezing-thawing procedure had no influence on the fertilization of testicular sperm, but epididymal frozen-thawed sperm had a higher fertilization rate. Immunofluorescence proved decreasing sperm quality in all groups compared with the control group. This difference is not manifested in other parameters (transfer, pregnancy, implantation rates). The spermatozoa with varied semen characteristics and good quality, also detected with specific antibodies, gave the best fertilization rates. The paternal effect is not proved in other parameters.

Chromosomal analysis of spermatozoa with normal-sized heads in two infertile patients with macrocephalic sperm head syndrome

To study the chromosomal content of spermatozoa that could be selected for intracytoplasmic sperm injection (ICSI) in cases of macrocephalic sperm head syndrome. Case report. Obstetrics, gynecology, urology, and reproductive biology departments. Two infertile patient candidates for ICSI presenting with total teratozoospermia (100%) with mainly large-headed spermatozoa (91% and 82%, respectively). Fluorescence in situ hybridization with X, Y, 18 centromeric probes on unselected spermatozoa (all migrated spermatozoa) and specifically on selected spermatozoa with normal-sized heads. Percentage of polyploid, diploid, aneuploid, and normal haploid spermatozoa, according to X, Y, 18 chromosome centromeric probes on selected spermatozoa (head size compatible with ICSI). All the nonselected spermatozoa were abnormal, diploid, or polyploid. The rate of normal ploidy (haploid cells) among the selected sperm population was 1 per 28 for patient 1 and 5 per 51 for patient 2. This very low proportion of normal haploid spermatozoa among selected spermatozoa contraindicated ICSI for the two patients. We suggest performance of this selection and analysis before including (or not), in an ICSI program, patients with macrocephalic sperm head syndrome, associated or not with preimplantation genetic diagnosis.

Influence of spermatogenic profile and meiotic abnormalities on reproductive outcome of infertile patients

Genetic aspects of male infertility and the possible risks of new assisted reproduction and their influence on the development of zygotes and children born after intracytoplasmic sperm injection (ICSI) need further research. These patients have an increased risk of diploidy, and disomies are frequent in their spermatozoa. Meiotic disorders are more common in testicular biopsies of patients with severe oligoasthenozoospermia. For these reasons, a detailed andrological study is absolutely mandatory before accepting a couple with these characteristics into an IVF–ICSI programme. When an andrological patient has plasma FSH values >10 IU/l and/or very low total motile sperm count <1 × 10 6, despite a normal karyotype, they clearly need a testicular biopsy and a meiotic study in order to rule out meiotic arrest or synaptic anomalies. Another important aspect to be considered is the possible benefit of applying preimplantation genetic diagnosis in these cases because they normally have a high percentage of chromosomally abnormal embryos, although in the present study this was not evident. All studies agree on the necessity of conducting follow-up studies in the population of children born after IVF–ICSI. In this way, it will be possible to find out if these infertile patients and their offspring have a higher risk of suffering epigenetic errors and imprinting disorders.

Toward single embryo transfer policy

Toward single embryo transfer policy

Comparison of Clinical Outcomes between the Gonadotropin-Releasing Hormone (GnRH) Antagonist, Cetrorenix, and the GnRH Agonist, Buserelin, during Controlled Ovarian Hyperstimulation in In Vitro Fertil

Objective: To compare clinical outcomes between the gonadotropin-releasing hormone (GnRH) antagonist, cetrorenix; and the GnRH agonist, buserelin, during controlled ovarian hyperstimulation (COH) in in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI) treatment. Material(s) and Method(s): This study was conducted on 57 cycles in 50 couples who used cetrorenix (group I) and on 68 cycles in 56 couples who used buserelin (group II) and were undergoing COH for IVF and ICSI programs. Recombinant FSH and human chorionic gonadotropin were administered for superovulation from cycle day 3 on an individual basis according to the ovarian response. Numbers of oocytes retrieved, oocytes fertilized, and cleaved embryos and pregnancy rates were compared between the two groups. Result(s): Numbers of oocytes and metaphase II oocytes retrieved in group II did not significantly differ from those in group I. The fertilization rate of metaphase II oocytes after insemination in group I (6.7 ± 0.6) was comparable with that of group II (7.3 ± 0.5). The clinical pregnancy rate and ongoing pregnancy rate were higher in group I (42.1% and 33.3%) than in group II (38.2% and 27.9%, respectively), but did not statistically significantly differ. No cancellation cycle occurred in either group. There was one instance of ovarian hyperstimulation syndrome (OHSS) observed in each group during the study. Conclusion(s): A multiple dose of the GnRH antagonist, cetrorenix, is as effective, efficient, and safe for COH as the long protocol of the GnRH agonist, buserelin, during IVF and ICSI treatment.

Apoptosis in human granulosa cells after induction of ovulation in women participating in an intracytoplasmic sperm injection program

Objectives: To investigate whether analysis of granulosa cell apoptosis can be useful in assessing follicular and oocyte maturation and the regulation of granulosa cell apoptosis by follicular fluid steroids in preovulatory follicles of stimulated women. Study design: Apoptosis in aspirated granulosa cells ( n=64) was measured using the Annexin V-affinity assay by flow cytometry. Follicular fluid steroids were determined by ELISA and RIA. Statistics were evaluated using the Levenne test, Student t-test and simple linear regression analysis. Results: No significant differences in the number and percentage of apoptotic granulosa cells per follicle were observed according to the maturity and fertilizability of the oocytes by intracytoplasmic sperm injection within these follicles. No correlations were found between levels of steroid hormones in follicular fluid and the number and proportion of granulosa cells undergoing apoptosis. Conclusions: The percentage of apoptosis in granulosa cells is not related to oocyte maturity and fertilizability by ICSI or to follicular quality in stimulated cycles of normal women. However, the possibility cannot be discarded that this parameter may be of importance at other phases of follicular development or in natural cycles when no treatment that influences follicular physiology is being applied.

Birth after intracytoplasmic injection of epididymal sperm from a man with congenital bilateral absence of the vas deferens who had a robertsonian translocation

Objective To present the first case of an infertile male with azoospermia related to a congenital bilateral absence of the vas deferens (CBAVD), in which mutations within the cystic fibrosis transmembrane conductance regulator (CFTR) gene coexist with a robertsonian translocation. Design Case report. Setting A university hospital. Patient(s) A 34-year-old male with a 2-year history of primary infertility. Intervention(s) Lymphocytic karyotype, polymerase chain reaction (PCR) and allele oligonucleotide-specific hybridization (ASO), microsurgical epididymal sperm aspiration (MESA), and intracytoplasmic sperm injection (ICSI). Main outcome measure(s) Physical examination and semen analysis. Result(s) Semen analyses revealed azoospermia and a well-recognized obstructive phenotype. Analysis of the CFTR gene revealed a compound heterozygosity for a 2184 del A + 2183 A → G mutation on one allele and the 5T variant within the polypyrimidine tract of intron 8 on the other allele. Cytogenetic analyses revealed a t(13;14)(q10;q10) robertsonian translocation in the same patient. Microsurgical epididymal sperm aspiration allowed retrieval of a million mature motile spermatozoa, excluding any spermatogenic impairment secondary to the genetic abnormalities found in this patient. Epididymal sperm was used for an intracytoplasmic sperm injection program, and a normal child was born at term. Conclusion(s) This case illustrates that two distinct genetic defects may coexist, with a variable effect on male fertility but with important implications for genetic counseling of the future pregnancy.

Binding of Zona Binding Inhibitory Factor-1 (ZIF-1) from Human Follicular Fluid on Spermatozoa

Previous studies showed that zona binding inhibitory factor-1 (ZIF-1) was the glycoprotein mainly responsible for the spermatozoa zona binding inhibitory activity of human follicular fluid. ZIF-1 has a number of properties similar to glycodelin-A. A binding kinetics experiment in the present study demonstrated the presence of two binding sites of ZIF-1 on human spermatozoa. These binding sites were saturable, reversible, and bound to (125)I-ZIF-1 in a time-, concentration-, and temperature-dependent manner. Glycodelin-A shared one common binding site with ZIF-1 on spermatozoa, and it could displace only 70% of the (125)I-ZIF-1 bound on human spermatozoa. ZIF-1 and glycodelin-A formed complexes with the soluble extract of human spermatozoa. Coincubation of solubilized zona pellucida proteins reduced the binding of ZIF-1 to two complexes of the extract, suggesting that the ZIF-1 binding sites and zona pellucida protein receptors on human spermatozoa were closely related. ZIF-1, but not glycodelin-A, significantly suppressed progesterone-induced acrosome reaction of human spermatozoa. The carbohydrate moieties derived from ZIF-1 reduced the binding of native ZIF-1 on human spermatozoa as well as the zona binding inhibitory activity of the glycoprotein, although the intensity of the effects are lower when compared with the native protein. These effects are not due to the action of the molecules on the motility, viability, and acrosomal status of the treated spermatozoa. Deglycosylated ZIF-1 had no inhibitory effect on both ZIF-1 binding and zona binding capacity of spermatozoa. We concluded that the carbohydrate part of ZIF-1 was critical for the functioning of the glycoprotein.

Correlation between human follicular diameter and oocyte outcomes in an ICSI program.

To determine the correlation between the follicular sizes and oocyte recovery, metaphase II oocyte recovery, fertilization rate and good embryo quality from mature and immature oocytes in an intracytoplasmic sperm injection (ICSI) program. 991 follicles obtained from 72 ICSI cycles were classified into three groups according to their diameters as measured by transvaginal ultrasound including group A (< 10 mm), group B (10-14 mm), and group C (> 14 mm). All obtained oocytes were classified according to their nuclear maturation: germinal vesicle (GV), metaphase I (MI) and metaphase II (MII). Mature oocytes underwent ICSI while immature oocytes were further cultured until maturity before ICSI was performed. The rates of fertilization and good quality embryos at day 3 were evaluated. A progressive and significant increase in the rates of oocyte recovery and MII oocyte recovery were observed from group A follicles compared to the other groups (p < 0.001). The fertilization rate of mature and in vitro matured oocytes, as well as the rate of good quality embryos showed a tendency to increase from group A to group C follicles, but not significantly. The corresponding fertilization rates were 78 and 55.3% (p < 0.001) for mature and in vitro matured oocytes, respectively. Collection of oocytes from small follicles, especially with a mean diameter less than 10 mm, and in vitro maturation of immature oocytes before fertilization may allow the total number of good quality and transferable embryos to be increased.

Sperm tail flexibility test: a simple test for selecting viable spermatozoa for intracytoplasmic sperm injection from semen samples without motile spermatozoa.

The objective was to describe the results of the injection of immotile spermatozoa with flexible tails when only immotile spermatozoa are present in the semen sample. A retrospective study was conducted to analyze the procedure results for 10 couples who participated in our intracytoplasmic sperm injection program. The sperm tail was considered flexible when it moved up and down independently of the head movement, and it was considered inflexible when the movement occurred together (tail plus head). The fertilization and pregnancy rate were analyzed. The normal fertilization rate (presence of 2 pronuclei) was 30.3% (40/132), and the abnormal fertilization rate (presence of less than or more than 2 pronuclei) was 6.81% (9/132). A total of 52 embryos were obtained with 9 transfer procedures performed (pregnancy rate: 11.12%). The sperm tail flexibility test (STFT) is an easy and cost-effective way for selecting viable immotile spermatozoa and can be used as an alternative method for determining the viability of spermatozoa. This test seems to be a simple and risk-free method when compared to the swelling test.

The effect of pentoxifylline (PF) in severe asthenozoospermia treatment on the results of ICSI program

Objective: Intracytoplasmic sperm injection (ICSI) is now widely used for male factor treatment in human IVF/ET program. Although poor sperm parameters including count, sperm motility and morphology has been reported not important in ICSI program, sperm motility is a prerequisite indicator to find viable spermatozoa in the treatment of severe asthenozoospermia. Pentoxifylline (PF) and other methylxanthines are known to influence sperm metabolism and motility. Because of its enhancing effect on sperm motility and its role in facilitating the acrosome reaction, the agent is now commonly used in human ICSI program to easily find motile spermatozoa. It has been postulated that PF, being a phosphodiesterase inhibitor, increase sperm kinetic parameters and the number of spermatozoa exhibiting hyperactivated motility by raising the intracellular content of cAMP, a molecule involved in the generation of sperm energy. The aim of study was to evaluate the effect of PF on the conventional ICSI program undergone in severe asthenozoospermia.

Lower sperm aneuploidy rate is associated with a higher pregnancy rate in unselected patients undergoing intracytoplasmic sperm injection

Objective: Infertile patients with oligo-astheno-teratozoospermia (OAT) and normal karyotype have a higher rate of sperm aneuploidy which is correlates with the severity of OAT. These patients frequently undergo intracytoplasmic sperm injection (ICSI) to father a child, raising concerns about the possible increased risk of chromosomal aneuploidies for the concepti, since a natural selection does not occur with this technique. Accordingly, few reports have shown that a high proportion of patients undergoing ICSI program have an increased sperm aneuploidy rate. Another important issue, addressed in this study, is to evaluate the impact of the abnormal sperm chromosome asset on ICSI outcome.Design: Forty-eight consecutive and unselected male partners of couples undergoing ICSI had sperm aneuploidy rate evaluated in the same sperm preparation used for ICSI and obtained by swim-up. They were divided into two groups whether their sperm aneuploidy rate was below (group A) or above (group B) the upper range (1.55%) found in 14 healthy normozoospermic men (control group). Their main ICSI outcome measures were compared.Materials/Methods: Sperm chromosome analysis was conducted by fluorescence in-situ hybridization (FISH), using alpha-centromeric DNA probes for chromosomes 8, 18 (two-color) and 12, X and Y (three-color FISH).Results: Twelve out of 48 patients (25%) had a total sperm aneuploidy rate below the cut-off value of the control group (median: 1.25%; range: 0.85–1.52%), whereas the remaining 36 (75%) patients had an elevated sperm aneuploidy rate (median: 3.25%; range: 1.64–23.60%). Group A patients’ age (median: 33 years; range: 23–40) was similar to that of group B patients’ (median 35 years; range: 25–52). Fertilization (93% vs. 85%) and cleavage (100% vs. 98%) rates resulted similar in both groups of patients. Patients of group A had a significantly higher clinical pregnancy (75% vs. 34%) and implantation (34% vs. 13%) rates compared to the patients of group B. Thirty-two percent of implanted embryos miscarried (preclinical and clinical abortion) in group B compared to the 11% of group A, but the difference did not reach the statistical significance. Other factors which may account for this difference, such as the age of the female partner, theirs infertility factor and the main sperm parameters, resulted to be similar in both groups of patients.Conclusions: This study confirmed that an elevated proportion of patients undergoing ICSI have an increased sperm aneuploidy rate and that cytogenetically abnormal spermatozoa have a negative impact on ICSI outcome.Supported by: A grant from MURST, Rome, Italy, 1999. Objective: Infertile patients with oligo-astheno-teratozoospermia (OAT) and normal karyotype have a higher rate of sperm aneuploidy which is correlates with the severity of OAT. These patients frequently undergo intracytoplasmic sperm injection (ICSI) to father a child, raising concerns about the possible increased risk of chromosomal aneuploidies for the concepti, since a natural selection does not occur with this technique. Accordingly, few reports have shown that a high proportion of patients undergoing ICSI program have an increased sperm aneuploidy rate. Another important issue, addressed in this study, is to evaluate the impact of the abnormal sperm chromosome asset on ICSI outcome. Design: Forty-eight consecutive and unselected male partners of couples undergoing ICSI had sperm aneuploidy rate evaluated in the same sperm preparation used for ICSI and obtained by swim-up. They were divided into two groups whether their sperm aneuploidy rate was below (group A) or above (group B) the upper range (1.55%) found in 14 healthy normozoospermic men (control group). Their main ICSI outcome measures were compared. Materials/Methods: Sperm chromosome analysis was conducted by fluorescence in-situ hybridization (FISH), using alpha-centromeric DNA probes for chromosomes 8, 18 (two-color) and 12, X and Y (three-color FISH). Results: Twelve out of 48 patients (25%) had a total sperm aneuploidy rate below the cut-off value of the control group (median: 1.25%; range: 0.85–1.52%), whereas the remaining 36 (75%) patients had an elevated sperm aneuploidy rate (median: 3.25%; range: 1.64–23.60%). Group A patients’ age (median: 33 years; range: 23–40) was similar to that of group B patients’ (median 35 years; range: 25–52). Fertilization (93% vs. 85%) and cleavage (100% vs. 98%) rates resulted similar in both groups of patients. Patients of group A had a significantly higher clinical pregnancy (75% vs. 34%) and implantation (34% vs. 13%) rates compared to the patients of group B. Thirty-two percent of implanted embryos miscarried (preclinical and clinical abortion) in group B compared to the 11% of group A, but the difference did not reach the statistical significance. Other factors which may account for this difference, such as the age of the female partner, theirs infertility factor and the main sperm parameters, resulted to be similar in both groups of patients. Conclusions: This study confirmed that an elevated proportion of patients undergoing ICSI have an increased sperm aneuploidy rate and that cytogenetically abnormal spermatozoa have a negative impact on ICSI outcome. Supported by: A grant from MURST, Rome, Italy, 1999.

Use of a highly sensitive quantitative telomerase assay in intracytoplasmic sperm injection programmes for the treatment of 47,XXY non-mosaic Klinefelter men.

We evaluated the role of the sensitive quantitative telomerase assay (SQTA) in the management of men with non-mosaic Klinefelter's syndrome (KS). Diagnostic testicular biopsy (DTB) was performed in 24 men with KS. A part of the DTB was stained and the remaining fragment was processed for the SQTA. After 3-18 months, a therapeutic testicular biopsy (TTB) was performed in the same testicle and the recovered specimens were processed to identify spermatozoa. Men with a SQTA outcome equal to 0.00 Units microg-1 protein (n = 7) demonstrated therapeutic testicular biopsy material that was negative for spermatogenic cells. In five men with a SQTA outcome of 8.11-38.03 Units microg-1, the most advanced germ cell was the spermatogonium/primary spermatocyte. In the remaining 12 men, the most advanced spermatogenic cell in the TTB was the spermatozoon. In these men, the SQTA outcome was equal to 25.76-92.68 Units microg-1 protein. Using 39.00 Units microg-1 protein as a cut-off value, the accuracy of the SQTA in identifying men positive for spermatozoa was 91.6%. It appears that the SQTA has a role for identifying non-mosaic KS men who have testicular spermatozoa.

Analysis of MUC-1 and integrin subunit expression in equine uterine epithelium and trophectoderm

Intracytoplasmic sperm injection (ICSI) is an important tool for equine embryo production in both clinical and research settings. In clinical ICSI programs, immature equine cumulus-oocyte complexes (COCs) are often collected at the mare's location and shipped to the ICSI laboratory. To simplify shipment and aid scheduling of subsequent procedures, COCs can be held overnight at room temperature (∼22 °C) before placement into maturation culture, with no detrimental effect on meiotic or developmental competence. A recent study indicated that it might be possible to hold COCs overnight at cold (∼4 °C) temperatures. If so, this might allow longer holding periods that would ease shipping requirements. In this study, we compared oocyte maturation rates, as well as cleavage and blastocyst rates after ICSI, for COCs held at either room or cold temperatures overnight before the onset of in vitro maturation. In Exp. 1, COCs were shipped overnight in a commercial embryo holding medium, ViGRO (Vg), in insulated containers designed to hold at either room temperature (RT, ∼22 °C) or cold temperatures (Cold, ∼7 °C). Subsequent rates of in vitro maturation, cleavage and blastocyst formation were significantly higher in the RT treatment (39%, 90% and 41%, respectively) than in the Cold treatment (23%, 60% and 17%, respectively, P < .05). In Exp. 2, we compared Vg medium with a second commercial embryo holding medium, SYNGRO (Sy). There was no significant difference between Vg and Sy groups in any evaluated parameter within either RT or Cold treatments. Within each medium group and for both media combined, the rates of in vitro maturation, cleavage and blastocyst formation were significantly higher in the RT treatment (42%, 81% and 42%, respectively for the combined media) than in the Cold treatment (29%, 54% and 10%, respectively for the combined media, P < .05). We conclude that shipment of immature equine COCs at cold temperatures (∼7 °C) is detrimental to subsequent in vitro maturation and embryo production.

General anesthesia versus monitored anesthesia care with remifentanil for assisted reproductive technologies: effect on pregnancy rate

Study Objectives: To compare the outcome of assisted reproductive technology procedures in women who undergo monitored anesthesia care (MAC) with remifentanil versus general anesthesia. Design: Retrospective data analysis. Setting: University hospital. Patients: 251 ASA physical status I and II women participating in an in vitro fertilization (IVF) or intracytoplasmic sperm injection (ICSI) program. Interventions: During the first phase of the study, all patients underwent general anesthesia induction with alfentanil, propofol, and nitrous oxide, which was maintained with isoflurane or propofol infusion. In the second phase of the study, all patients received a standardized MAC technique with a remifentanil infusion; local anesthetics were not used. Measurements: The primary endpoint was pregnancy rate per transfer. The number of oocytes collected, fertilized, and cleaved was recorded, as was the number of oocytes transferred. Main Results: Patients who underwent MAC had a greater pregnancy rate with IVF (28.2 vs. 16.3%), with ICSI (32.2% vs. 18.8%), and overall (30.6% vs. 17.9%). Conclusions: Pregnancy rates in women undergoing transvaginal oocyte retrieval for assisted reproductive technologies were significantly higher with a remifentanil-based MAC technique than with a general anesthetic technique.