Laparoscopic Follicular Aspiration Research Articles

Follicular response and oocyte production following variations in ovarian stimulation in goats

In this study, goats were subjected to ovarian stimulation protocols to evaluate possible differences in ovarian follicular responses and oocyte production. Two experiments were conducted to assess the effects of hormonal protocol duration (seven or twelve days) and number of follicle stimulating hormone (FSH) applications (one or five doses). All animals received intravaginal sponges saturated with 60 mg medroxyprogesterone acetate and an application of 125 μg cloprostenol 72 h before the sponges were removed. For ovarian stimulation, 120 mg FSH was applied in a single dose 36 h before laparoscopic follicular aspiration (LOPU) or in five doses (30, 30, 20, 20, and 20 mg) at 12 h intervals, with the last dose applied 36 h before LOPU. In the first experimental phase, ultrasonography was performed to monitor follicular number and diameter, and in the second phase, the animals received LOPU to count the follicles and cumulus-oocyte complexes (COCs) and for morphological classification. There was no significant effect (P > 0.05) of any variable or combination of variables on follicle number on Day (D) 0 or D3/D8 (day of LOPU). However, evaluation at D6/D11 revealed an effect (P < 0.05) from the protocol duration with the highest number of small follicles resulting from the short protocol. There was also an effect (P < 0.05) of FSH dose number on the resulting number of medium and large follicles, with more medium follicles recovered after a single dose and more large follicles after multiple doses. There was no effect of any variable (P > 0.05) on the diameters of the large, medium, and small follicles, except for the D4/D9 evaluation, which showed a combination effect for large follicles (P < 0.05). In the second phase, no variable had an effect on the number of follicles visualized or aspirated, number of COCs recovered, recovery rate, morphological quality of COCs in grades 1, 2, 3, and 4, or rate of viable COCs. Thus, all four protocols efficiently stimulated the ovarian response and oocyte production of goats, although the short protocol (7 days) with a single dose of FSH was most efficacious based upon the greater number of medium-sized follicles available for aspiration, the shorter time, and greater practicality of execution.

116 IMPROVED SURVIVAL BY CRYOPRESERVING RHESUS MACAQUE (MACACA MULATTA) SPERMATOZOA WITH DIRECTIONAL FREEZING TECHNIQUE

A significant increase in nonhuman primate models of human diseases will be expected in the near future since the successes in production of genetically engineered rhesus monkey models of human diseases. Sperm banking can provide an effective way to preserve valuable genetic resources. Our objective was to (1) develop a protocol using directional freezing technique (DFT) for rhesus monkey spermatozoa cryopreservation, which allows precise control of the velocity and the morphology of the ice-front propagation by transferring the tubes loaded with 2 mL sperm samples at a controllable velocity through two separate chambers with controllable temperature settings, and (2) achieve survival rate that was higher than that achieved with conventional freezing technique (CFT), by which sperm samples were cryopreserved in 0.25 mL straws with liquid nitrogen vapor in a styrofoam box. Sperm motility, acrosomal integrity, and in vitro fertilization (IVF) assay were used to assess the function of frozen-thawed spermatozoa. Data were analyzed by ANOVA and Fisher protected LSD test. Experiment 1 was aimed at optimizing the cooling rate using DFT. Tubes were frozen using the multi-thermal gradient freezing device (MTG 516, Harmony CryoCareTM, IMT Ltd.) at fast (16°C/min), medium (12°C/min), and slow (7°C/min) cooling rates, which corresponded to the transferring velocities (2.5, 1.5, and 0.5 mm s-1, respectively). The results showed that spermatozoa frozen at fast and medium cooling rates showed significantly higher frozen-thawed motility than those frozen at slow cooling rate (61% and 59% v. 50%, P &lt; 0.05). However, no difference was observed on sperm acrosomal integrity among the experimental groups (84, 80, and 78%, respectively, P &gt; 0.05). The purposes of Experiment 2 were determined to examine if using DFT at the optimized cooling rate (12°C/min) can improve the cryo-survival of rhesus monkey spermatozoa compared with CFT. Our results showed that spermatozoa cryopreserved by using DFT achieved significantly higher frozen-thawed sperm motility that those cryopreserved by using CFT (64 v. 54%, P &lt; 0.05). However, no difference was observed on acrosomal integrity between spermatozoa cryopreserved by DFT and CFT (84 and 83%, respectively; P &gt; 0.05). The function of spermatozoa cryopreserved by using DFT was further evaluated by IVF. Females were treated with rhFSH twice-daily for 8 days after the onset of menses and following a treatment of hCG injection on Day 9. Cumulus-oocyte complexes were collected by laparoscopic follicular aspiration 32 h later. Of the inseminated oocytes, 79% were fertilized and 90 and 53% of the resulting zygotes developed into 2-cell and blastocysts, respectively. The fertilization rate was lower and the blastocyst rate was slightly higher than our previous report when fresh spermatozoa were used for IVF (94 and 52%, respectively). Our results indicate that spermatozoa of rhesus monkeys can be effectively cryopreserved using DFT in large volume. This finding provided a new and effective way for genetics preservation purposes in this important species.

Evaluation of the vervet (Clorocebus aethiops) as a model for the assisted reproductive technologies

The vervet monkey was evaluated as a primate model for use in assisted reproductive technologies (ARTs). Eight adult female vervets were hormonally monitored for their potential use as egg donors and those six females displaying regular menstrual cycles were subjected to controlled ovarian stimulation with recombinant human gonadotropins. Three animals failed to respond while laparoscopic follicular aspiration was performed on the other three females at 27-30 h post-human chorionic gonadotropin administration. A total of 62, 40, and 18 oocytes was recovered from these three animals of which 30, 20, and 4, respectively, matured to the metaphase II stage and were subsequently inseminated using intracytoplasmic sperm injection. An average of 40+/-15% (SEM) of the inseminated oocytes were fertilized based on pronucleus formation and timely cleavage. One embryo from each of the two stimulated females developed into expanded blastocysts. Two adult male vervets were assessed as sperm donors. Neither adjusted well to the restraint and collection procedure required for penile electroejaculation. Samples collected via rectal electroejaculation were very low in sperm motility and concentration; however, cauda epididymal aspirations from one male yielded an adequate concentration of motile sperm. These results emphasize the need to establish species-specific ovarian stimulation protocols and semen collection techniques if vervets are to be considered for basic and applied (ARTs) research on primate gametes or embryos.

Oestradiol concentration as a predictor of ovarian response in FSH stimulated ewe-lambs

We investigated the prediction of ovarian response using oestradiol determination, in 37 gonadotrophin stimulated Karagouniko ewe-lambs. Ovarian stimulation was induced by serial FSH administrations, and laparoscopic follicular aspiration (OPU) was conducted 12 h after the last FSH injection. Oestradiol concentration was assessed in six blood samples collected prior to each FSH injection and in one sample collected prior to follicular aspiration. According to ovarian response, ewe-lambs were allotted in three groups: good, L1 ( n = 17); moderate, L2 ( n = 10); and poor, L3 ( n = 10). Based on the data obtained from 28 (75%) randomly selected animals, a statistical model was designed and tested on the remaining nine lambs for its ability to predict the probability of good ovarian response. From the 2nd sample, oestradiol concentration was constantly higher in group L1 in comparison with L3 lambs (all p-values for the contrasts were ≤0.02), while this difference between L1 and L2 lambs was significant only in the 6th and 7th sample (both p < 0.005). Using as criterion the oestradiol concentration of the 6th sample, the statistical model predicted all lambs that did not belong to group L1 and three of four lambs that belonged to group L1. Our results indicate that the moderate- and poor-donors could be safely predicted on the basis of oestradiol concentration 12 h prior to the scheduled follicular aspiration. Moreover, poor-donors could be identified—and rejected from further manipulations—on the basis of their inability to exhibit increased oestradiol levels in response to gonadotrophin stimulus.

Ovum Pick‐up in Sheep: a Comparison between Different Aspiration Devices for Optimal Oocyte Retrieval

In vivo ovum pick-up (OPU) in sheep may be improved with a proper choice of aspiration elements (needle and tubing) and aspiration vacuum pressure. In the present study, two experiments were carried out. In Expt 1, visible follicles in ovaries of slaughtered ewes (treated separately according to their diameters: small<3 mm, medium 3-5 mm and large>5 mm) were aspirated using different combinations of the three studied factors such as aspiration flow rate (10, 20, 30, 40 and 50 ml water/min), needle gauge (18 and 20 G) and tubing inner diameter (1, 2 or 3 mm internal diameter). In Expt 2, a study with two 18 G needles of different lengths (18 G: 82 mm; 18 GL: 600 mm) was carried out, using ovaries obtained post-mortem, and performing in vivo laparoscopic follicular aspiration on ewes. We considered good quality oocytes as those with both complete compact cumulus and a homogeneous cytoplasm. Recovery rate, proportion of good quality oocytes (good quality oocytes/100 oocytes recovered) and overall efficiency (good quality oocytes/100 follicles aspirated) were noted. In Expt 1, aspiration flow rate affect remarkable proportion of good quality oocytes (69.5%, 50.5%, 44.8%, 36.5% and 28.3% for flows from 10 to 50 ml/min respectively, p<0.05). Needle gauge did not affect aspiration device efficiency. Thin and intermediate tubings were more effective (overall efficiency rates: 34.9%, 32.3% and 28.1% for 1, 2 and 3 mm respectively, p<0.05). Follicle size did not affect recovery rate, but proportion of good quality oocytes was higher for large (77.9%) and medium (64.4%) follicles (p<0.05). Finally, some combinations of the aspiration device showed greater effectiveness. In Expt 2, needle length did not influence recovery rate, but good quality oocytes rate was significantly modified both post-mortem and in vivo (good quality rate for 18 G vs 18 GL needles: 69.5% vs 47.7% and 58.1% vs 25.4%, post-mortem and in vivo respectively, p<0.05). We conclude that low-aspiration flow rates (10 and 20 ml/min) with thin or intermediate tubings (1 and 2 mm), and any short needle (18 G or 20 G) are the most adequate aspiration factors for OPU in sheep.

160EFFECTS OF EXTRACELLULAR ION CONCENTRATIONS ON IN VITRO DEVELOPMENT OF DOMESTIC CAT IVF EMBRYOS

Domestic cat embryos typically are cultured in media formulated for somatic cells or embryos from rodents or livestock species. Under these conditions, blastocyst development has been inconsistent and delayed relative to embryos grown in vivo, and embryo viability following transfer has been low. Our goal is to systematically define the culture requirements of the feline embryo to improve embryo development and viability. The objective of this study was to determine the ionic (NaCl, KCl, KH2PO4, and CaCl2:MgSO4) preferences of domestic cat IVF embryos. Anestral female cats were injected (i.m.) with 150IU eCG followed 84h later by 100IUhCG. Oocytes were recovered via laparoscopic follicular aspiration approximately 24h post-hCG injection (Day 0). Semen was collected from one of two males by means of an artificial vagina and washed once in HEPES-buffered IVF medium. Mature cumulus-oocyte complexes were co-incubated with 2.5–5×105 motile sperm mL−1 in IVF medium (100mM NaCl, 4.0mM KCl, 1.0mM KH2 PO4, 2.0mM CaCl2, 1.0mM MgSO4-7H2O, 25.0mM NaHCO3, 3.0mM glucose, 0.1mM pyruvate, 6.0mM L-lactate, 1.0mM glutamine, 0.1mM taurine, 1×MEM nonessential amino acids, 50μgmL−1 gentamicin, and 4.0mgmL−1 BSA) for 19 to 22h in 6% CO2 in air (38.7°C). Cumulus cells were removed and embryos cultured (8–11 embryos/50μL drop; 6% CO2, 5% O2, 89% N2, 38.7°C) in media containing 100.0 or 120.0mM NaCl, 4.0 or 8.0mM KCl, 0.25 or 1.0mM KH2PO4, and 1.0mM:2.0mM or 2.0mM:1.0mM CaCl2:MgSO4 (2×2×2×2 factorial design). The remaining components of the culture medium were identical to the IVF medium (but w/o gentamicin). Development to the blastocyst stage by Day 6, metabolism (glycolysis and pyruvate) of each blastocyst, and final cell number (Hoechst 33342 staining) of all embryos were evaluated. Final cell number of cleaved embryos and development to the blastocyst stage were analyzed using analysis of variance in the GLIMMIX macro of SAS. A total of 236 oocytes were inseminated, yielding 128 cleaved embryos (54%), including 6 blastocysts (4.7% of cleaved embryos). Cell number was not (P&amp;gt;0.05) affected by NaCl, KCl, or KH2PO4 concentrations, but tended (P=0.057) to be higher after culture in 2.0mM:1.0mM CaCl2:MgSO4. Treatments did not significantly affect (P&amp;gt;0.05) development to the blastocyst stage, but numerically more blastocysts were produced in 100.0mM NaCl (4/6), 8.0mM KCl (5/6), or 1.0mM KH2PO4 (5/6). Both CaCl2:MgSO4 ratios resulted in 3 blastocysts. Blastocysts contained 61.08±5.1 (mean±SEM, n=6) cells and actively metabolized glucose (glycolysis, 3.7±0.8pmol/embryo/3h or 0.06±0.01pmol/cell/3h) and pyruvate (0.75±0.27pmol/embryo/3h or 0.013±0.005pmol/cell/3h). These results suggest that the ionic composition of culture media influences the in vitro development of cat IVF embryos. (Supported by NIH grant RR15388.)

Caprine somatic cell nuclear transfer using in vivo matured oocytes collected by laparoscopic follicular aspiration

ABSTRACT In vivo matured oocytes collected by laparoscopic follicular aspiration (LFA) from hormone treated female goats were used as recipient ooplasts for somatic cell nuclear transfer (SCNT). Japanese native (Shiba) goats were used as donor females and some donor females were used repeatedly (two or three times) at intervals of a few months. To induce synchronization of estrus, a sponge containing 0.5 g of progesterone was inserted into the vagina of each goat for 14 days. These animals were also treated with follicle stimulating hormone (FSH) in a series of 8 injections over 4 days. The first FSH injection was administered on the morning of day 9 of sponge insertion. On the morning of day 13, 50 µg of gonadotropin‐releasing hormone (GnRH) was injected into each animal. Twenty‐nine hours after GnRH injection, LFA was performed. After removal of cumulus cells, collected oocytes with the first polar body were selected and enucleated for nuclear transfer. Anterior pituitary cells isolated from an adult male Shiba goat were transfected with a DNA fragment containing the enhanced green flourescent protein gene and the puromycin resistance gene. A single donor cell was inserted into the perivitelline space of each enucleated oocyte and fusion was induced with one electric pulse of 20 V for 10 µs. The SCNT goat eggs were cultured in chemically defined medium at 38.5°C in 5% CO2, 5% O2, 90% N2 for 9 days. By LFA, 396 oocytes were collected from a total of 30 females. After removal of cumulus cells, 64% of them extruded the first polar body. The percentage of SCNT goat eggs produced using in vivo matured oocytes which developed to the blastocyst stage (20–21%) was significantly higher (P &lt; 0.05) than that produced with in vitro matured oocytes (3–8%).

Efficacy of porcine gonadotropins for repeated stimulation of ovarian activity for oocyte retrieval and in vitro embryo production and cryopreservation in Siberian tigers (Panthera tigris altaica).

A comparison of the amino acid sequences demonstrated that Siberian tiger gonadotropins are more homologous with those of porcine than any other commercially available preparation. The present study measured the efficacy of repeated ovarian stimulation with purified porcine gonadotropins on the follicular, hormonal, and immunogenic responses in Siberian tigers as well as on the ability of oocytes retrieved by laparoscopic follicular aspiration to fertilize and cleave in vitro. Controlled rate and vitrification cryopreservation methods were also compared for their ability to support ongoing cleavage following thawing of presumptive 2- to 4-cell tiger embryos generated in vitro. Vitrification supported continued embryonic cleavage in vitro while controlled rate freezing did not. Stereological microscopy indicated an excellent ovarian response with the recovery of quality cumulus-oocyte complexes that apparently fertilized and cleaved in vitro. However, ultrastructural and physiological examination revealed abnormal and unnatural responses such as the failure of some cumulus-oocyte complexes to reach maturity and progestagen levels to approach normalcy. At the same time, analyses of blood for antibodies failed to detect an immune reaction to these foreign gonadotropins in an assay that tested positive for the chorionic gonadotropin-stimulated domestic cat. Together, these observations suggest that porcine gonadotropins may be effective for the ovarian stimulation of tigers but that some modifications to administration protocols are needed to produce a more natural response.

Successful in vitro production of embryos in the red deer ( [formula omitted][formula omitted]) and the sika deer ( [formula omitted][formula omitted

The aim of our study was to define the conditions for IVM and IVF of oocytes in 2 common deer species as models for endangered related subspecies. Immature oocytes were recovered during the breeding season from postmortem ovaries (red deer) or by repeated laparoscopic follicular aspiration (sika deer). Oocytes were cultured for 24 h in IVM medium supplemented with EGF or FSH and follicular fluid. Stag semen was collected by electroejaculation (both species) or by epididymal flushing (red deer) and cryopreserved. For IVF, oocytes were exposed to different concentrations of thawed spermatozoa in a modified Tyrode albumin lactate pyruvate medium supplemented with 20% (v / v) estrus sheep serum for 18 h. After IVF, presumptive zygotes were allowed to develop in vitro for 7 days in synthetic oviduct fluid (SOF) supplemented with fetal calf serum (10%, v / v). In both species, the presence of ovine FSH and follicular fluid improved the in vitro maturation rate. In the sika deer, the optimal sperm concentration for IVF was 10 6 / mL and some fertilized oocytes reached the early morula stage (20 to 25 cells). In the red deer, after IVF with ejaculated or epididymal spermatozoa (2.0 x 10 6 / mL), 20% of zygotes developed to the blastocyst stage (50 to 80 cells).

Efficiency of laparoscopic follicular aspiration in sheep

Efficiency of laparoscopic follicular aspiration in sheep

Hyperexpression of epidermal growth factor receptors in granulosa cells from women with polycystic ovary syndrome

Objective: To compare the localization and quantitation of epidermal growth factor (EGF) receptor in granulosa cells from women with normal and polycystic ovaries. Design: Controlled, comparative study. Setting: Academic research laboratory. Patient(s): Forty-two women with normal or polycystic ovaries who attended our facility for the recovery of their immature or mature oocytes or for therapeutic purposes. Intervention(s): Patients underwent intravaginal ultrasound-guided oocyte retrieval or laparoscopic follicular aspiration with or without prior stimulation. Main Outcome Measure(s): Quantitation of EGF receptor in granulosa cells. Result(s): Granulosa cells from polycystic ovaries expressed significantly higher levels of EGF receptor than granulosa cells from normal ovaries. In contrast with patients who were treated with clomiphene citrate, those who were treated with gonadotropins showed low levels of the receptor. However, the levels of the receptor in granulosa cells were not correlated with circulating levels of LH, FSH, progesterone, or E 2. Immunolabeling of EGF receptor was confined to the cell membrane of granulosa cells. This receptor was fully functional, mediating the ligand-induced inhibition of E 2 production in culture. Conclusion(s): These results provide further evidence supporting a possible role of EGF/transforming growth factor-α in the aberration of ovarian function in polycystic ovary syndrome.

Repeated laparoscopic follicular aspiration in lambs

Repeated laparoscopic follicular aspiration in lambs

Follicular fluid α-fetoprotein, carcinoembryonic antigen, and CA-125 levels in relation to in vitro fertilization and gonadotropin and steroid hormone concentrations

To investigate the possible role of alpha-fetoprotein (AFP), carcinoembryonic antigen (CEA), and CA-125 in the ovarian follicle. alpha-Fetoprotein, CEA, and CA-125 were measured in human follicular fluid (FF) and correlated to IVF outcome and gonadotropin and gonadal steroid hormones. The In Vitro Fertilization Program at Virgen de las Nieves Hospital, Granada, Spain. Thirty-six FF from 12 infertile women with irreparable tubal damage who underwent treatment for ovarian stimulation with clomiphene citrate, hMG, and hCG. Laparoscopic follicular aspiration followed 48 hours later by ET. Serum and FF AFP, CEA, and CA-125 levels. alpha-Fetoprotein, CEA, and CA-125 were detected in all samples of serum and FF. There were no significant differences between the serum and FF levels. Follicular fluid AFP, CEA, and CA-125 of fertilized oocytes were similar to those in the unfertilized oocytes group. There were no significant correlations between the FF AFP, CEA, and CA 125 levels and gonadotropin and gonadal steroid hormone levels. alpha-Fetoprotein, CEA, and CA-125 are present in FF after ovarian stimulation, but the mean intrafollicular levels do not differ significantly regardless of the outcome of oocyte IVF.

Changes in follicular fluid gas and pH during carbon dioxide pneumoperitoneum for laparoscopic aspiration and their effect on human oocyte fertilizability

To determine changes in follicular fluid (FF) gases and acidity (pH) during either ultrasound (US)-guided or laparoscopic aspiration under carbon dioxide (CO2) pneumoperitoneum and relate the findings to the outcome of oocyte fertilization in vitro. A prospective study carried out over a limited period of 1 month during which test FF samples were obtained without direct exposure to CO2 or air. Within the assisted pregnancy program setting at the Human Reproductive Biology Unit at the Soliman Fakeeh Hospital. Infertile women undergoing either laparoscopic or US guided follicular aspiration for the purpose of in vitro fertilization treatment. Laparoscopic follicular aspiration under 100% CO2 pneumoperitoneum or transurethral US-guided follicular aspiration. Mean FF carbon dioxide partial pressure (pCO2) and oxygen partial pressure (pO2) were significantly higher in those aspirated at laparoscopy compared with those obtained by US-directed aspiration. The mean FF pH in the laparoscopic group was as a consequence significantly lower than in the US group. Oocyte fertilizability was significantly reduced in the laparoscopic aspiration group compared with the US group. This was more pronounced when the fertilization rate of the last recovered oocytes from the groups were compared. No difference was observed in the post fertilization embryonic development between the two groups. One hundred percent pneumoperitoneum leads to changes in FF that adversely affect oocyte fertilizability in a time-dependent manner. The use of US-directed procedure is advocated for all patients undergoing follicular aspiration in assisted pregnancy program. Where this is not feasible, the duration of CO2 pneumoperitoneum should be minimized.

Pregnancies and live birth from in vitro fertilization of calf oocytes collected by laparoscopic follicular aspiration

Oocytes were recovered by laparoscopic aspiration from 3- to 8-week-old calves treated with follicle-stimulating hormone (FSH) followed by human chorionic gonadotropin (hCG) to induce follicular growth and oocyte maturation in vivo. Most of the recovered oocytes either had resumed meiotic maturation at the time of aspiration or were competent to undergo maturation during subsequent culture in vitro. Oocytes matured in vivo following FSH and hCG treatment underwent in vitro fertilization (70%) at rates not significantly different from those of control oocytes recovered from adult cow ovaries at abattoirs and matured in vitro (75%). Calf oocytes that were immature at aspiration exhibited lower fertilization rates after in vitro maturation (36%) but their rate of development to morulae and blastocysts did not differ from that of mature oocytes at aspiration. A total of 91% of the zygotes produced from calf oocytes developed to morula and 27% to blastocyst stages during 6 days of culture. The proportion developing to morulae was significantly higher (P<0.05) than that observed for zygotes resulting from in vitro maturation and fertilization of oocytes recovered from cow ovaries obtained at an abattoir and processed concomitantly (59% to morulae and 18% to blastocysts). Morulae or blastocysts developed from oocytes from 5 to 6-week-old calves, when transferred to synchronized recipient heifers, resulted in 2 confirmed pregnancies, one of which produced a single full-term live calf. The ability to produce embryos from oocytes recovered from newborn or prepubertal calves offers the potential for markedly reducing the generation interval in cattle, thereby substantially accelerating the rate of genetic gain that can be achieved through embryo transfer.

Ovarian stimulation with pure follicle-stimulating hormone/human menopausal gonadotropin and improved laparoscopic aspiration needles influence the success of an in vitro fertilization program

The effect of a combined pure follicle-stimulating hormone/human menopausal gonadotropin (pFSH/hMG) ovarian stimulation regimen and modified, sharpened laparoscopic follicular aspiration needles on the number of oocytes retrieved and the oocyte/follicle ratio in 43 consecutive cycles of in vitro fertilization (IVF) were retrospectively compared with 99 consecutive preceeding cycles stimulated with hMG alone and captured with aspiration needles that had never been sharpened. A modified laparoscopic follicular aspiration needle is described. Purified FSH/hMG ovarian stimulation significantly improved the mean serum estradiol levels, number of preovulatory follicles, and, therefore, the total number of oocytes recovered per cycle. The mean ratios of oocytes recovered per preovulatory follicle documented on ultrasound, and per aspirated follicle, increased significantly using sharpened needles. Both modifications improved the success rate of our IVF program.

Comparison of ultrasonographic transurethral and laparoscopic guided oocytes retrieval

Fifty-eight patients who underwent in vitro fertilization (IVF) treatment were divided into two groups. In 28 patients, ultrasonically guided transurethral follicular aspiration was performed under light general anesthesia. In 30 patients, laparoscopic follicular aspiration was performed under general anesthesia. The mean age for both groups was similar. Follicular recruitment was achieved with human menopausal gonadotropin (hMG) or a combination of clomiphene citrate and hMG. In the ultrasonically guided aspiration group, 128 follicles were aspirated, with a 64% recovery rate (83 oocytes) and an 88.1% embryo transfer (ET) rate (67 embryos). Three (10.7%) intrauterine pregnancies were established in this group. In the laparoscopic group, 153 follicles were aspirated, with a 68.6% recovery rate (105 oocytes) and an 86.3% ET rate (82 embryos), with 4 (13.3%) intrauterine pregnancies established. Three patients had both procedures done; however, none conceived. There were no statistically significant differences between the two groups. Ultrasonically guided transurethral follicular aspiration should be considered as an alternative route for oocyte retrieval, especially when laparoscopy is contraindicated or when the ovaries are not accessible.

The effect of laparoscopic follicular aspiration on ovarian steroidogenesis during the early preimplantation period

Seventeen patients were studied for the first 72 hours of the luteal phase after spontaneous ovulation or follicle aspiration. Nine patients (group I) underwent follicle aspiration after clomiphene citrate (CC) administration, three women were studied after CC for ovulation induction (group II), and five spontaneous ovulating subjects served as controls (group III). Serum progesterone (P) concentrations were significantly elevated in stimulated cycles with or without follicle aspiration, compared with subjects. Moreover, when aspirated subjects were compared with nonaspirated stimulated subjects, a significant difference in P concentration was identified. It can be concluded that with follicle aspiration, P decreases can be observed, but with multiple follicle development, inadequate P levels are not seen in these stimulated subjects. On the contrary, with such elevated P levels, endometrial advancement may play a role in poor implantation success with human in vitro fertilization procedures.

In vitro fertilization in a community hospital.

Blodgett Memorial Medical Center began developing an IVF program in September of 1982. It took 10 months to establish animal and human IVF laboratories and an andrology laboratory. All personnel were recruited from within the hospital. From August 1983 through March 1985, 49 patients went through 58 complete IVF laparoscopy cycles. hMG and hCG were used to stimulate folliculogenesis and ovum maturation. An average of 5.2 oocytes was obtained by laparoscopic follicular aspiration. Eighty-four percent were either mature or intermediate oocytes. The fertilization rate per oocyte was 67%. Eighty percent underwent cleavage. Forty-seven patients (81%) received 1 or more embryos, with an average of 2.8 embryos each. There have been 12 pregnancies, including 3 abortions and 3 multiple pregnancies. Eight healthy infants have been born to date. The pregnancy rate per laparoscopy cycle was 21% and per transfer cycle was 26%. As a result of the IVF program, other services for infertile couples have been developed.

Ultrastructure of Baboon Zygotes Produced By Microinjection of Spermatozoa

Five baboons were treated during seven menstrual cycles with 5.0 mg of FSH-P for five days, starting on either day 3 or day 5 of the cycle. On day 5 of the treatment, the ovaries were examined by laparoscopy to evaluate follicular development. All animals exhibited multiple preovulatory follicles and at that time 100 mg GnRH was administered intramuscularly to induce LH release. Between 24 and 30 hours after injection of GnRH, laparoscopic follicular aspiration was used to collect oocytes. These were matured in vitro (determined by extrusion of the first polar body) and fertilized by microinjection with frozen-thawed baboon spermatozoa. Male and female pronuclei were observed in 32% of the resulting zygotes within 24 hours. These zygotes were compared to a zygote in the same stage that had been fertilized in vivo and obtained by laparoscopy and flushing of the oviduct.