The effect of sperm parameters on the outcome of intracytoplasmic sperm injection

310

Citations from the literature /International Journal of Gynecology & Obstetrics 53 (1996) 305-316

fertile semen in sperm-zona binding was examined under hemizona assay conditions. One droplet of a suspension of spermatozoawas exposed to sperm proteins and then tested for zona binding, while a parallel semen suspension droplet incubated with culture medium served as a control. The reliability of the test was increased by relating the number of spermatozoa bound to each inseminated hemizona to the surface area of the hemizona and expressedas the binding index. For spermatozoa incubated with extracted proteins, the binding index was greater than (jr = 0.001) that of: controls (125.2 l 45.1 vs. 63.6 * 29.2, respectively). As a first control, two other protein sources (fetal calf serum and human follicular fluid) were tested in the hemizona assay. No significant differenceswere found in zona binding for other proteinexposedspermatozoacompared with controls. As a secondand reverse control, exposure of one hemizona to sperm proteins before insemination with untreated spermatozoa induced a marked decmase(p = 0.0003)in sperm binding, compared with that of the matched hemizona not exposed to sperm proteins (control) (3.4 i 1.4 vs. 74.5 f 6.8, respectively). Taken together, these findings confirm the involvement of extracted sperm proteins in sperm-zona interactions. Therefore, in the easesin which fertilization in vitro fails because of a lack of sperm-zona binding, incubation of deficient spermatozoa with proteins extracted from spermatozoa of fertile ejaculates should restore their ability to interact with the oocyte and, thus, should enhance the prognosis for in vitro fertilization. -eata-ef p=!tP=Ybym~~~ intmtdm~t&nwIth~ncoveredbyamadiBed H~~fmmap8t&ntwitbrotmgradeejaenIotloo Ranieri D.M.; Siionetti S.; Vicino M.; Cormio L.; Selvaggi L. ITA FERTIL STERIL 19956415(1039-1042) Objective: To improve the quality of the sperm recovered from the bladder in a patient with retrograde ejaculation who already had failed to conceive after several attempts at IUI with sperm recovered by conventional techniques. Setting: University Hospital. Patients: A couple with male infertility due to retrograde ejaculation causedby the Zielke operation, a spinal fuation procedure performed to correct severe.kyphoscoliosis. Intervention: Superovulation and IUI of sperm recovered from the bladder using a modified Hotchkiss procedure involving the introduction into the bladder of Earle’s balanced salt solution (EBSS) buffered with Hepes in sufficient quantity to bring the urinary pH and osmolarity to those of fresh ejaculate. Main Outcome Measures: Urine pH and osmolarity at baseline and after dilution with EBSS buffered with HEPES. Concentration, motility, and progression scoreof the sperm recoveredfrom the bladder. Results: Good sperm samples were achieved. Pregnancy was established when IUI was performed in association with superovulation induction. Conclusions: Determination of urine pH and osmolarity appears to be a useful method for choosing the ideal sperm recovery procedure. The modified Hotchkiss procedure describedseemsto be a promising altemative method for recovering sperm for artificial insemination.

slrceasfpl~ontcomeaftcrcryoprcservatieoofaBfre-sb embryos with dmqwat trader iota an mwtimdated cycle

Frederick J.L.; Ord T.; Kettel L.M.; Stone S.C.; Balmaceda J.P.; Asch R.H. USA FERTIL STERIL 199564/5 (987-990) Objective: To assessthe pregnancy outcome of freezing and storing all fresh embryos produced in a stimulated IVF cycle and replacing them in a subsequent nongonadotropinstimulated cycle. Design: Retrospective study. Setting: University-associated assisted reproductive technology program. Patients: We studied 36 patients (age range 23 to 44 years) who underwent cryopreservation of all fresh embryos in a controlled ovarian hyperstimulation (COH) cycle because of either the risk of severe ovarian hyperstimulation (24 patients, group 1) or the presenceof an endometrial lining c 8 mm in thickness (12 patients, group 2). Five hundred fifty-five embryos were gemrated for replacement in 63 cycles. All embryos were cryopreserved in 1.5 M propanediol at the pronuclear or hvocell stage, and 264 embryos subsequently were transferred into a hormone replacement cycle (79%) or natural ovulatory cycle (38%).The average number of embryos transferred per patient was 4.2. Results: Twenty-one clinical pregnancieswere achieved, giving a pregnancy rate (PR) of 58.3% per patient (33.3% per cycle). The live birth rate was 56% per patient (28.6% per cycle). The implantation rate was 9.1%. Groups 1 and 2 had a similar PR per patient (58.3%). With 208 cryopreserved embryos remaining and considering the 33.3% PR per cycle, we expect the overall extrapolated PR to be 63.9%. Conclusions: This is the first seriesshowing that freezing and storing all fresh embryos produced in a stimulated IVF cycle and replacing them in a subsequentnongonadotropin-stimulated cycle results in successfulPRs. These results underlie the importance of a successfulcryopreservation program in IVF and could be a possible approach to overcoming the alleged adverseeffects of COH on the endometrium, thereby improving the chances of pregnancy when numerous embryos are obtained simultaneously. llleeffectofspermpammeterswtbetmk!omeofintraCYw-kfJP-~ieetioll

Mansour R.T.; Abouighar M.A.; Serour G.I.; Amin Y.M.; Ramzi A.M. EG FERTIL STERIL 199564/5 (982-986) Objective: To investigate the infiuence of sperm parameters on the fertilization and pregnancy rates in intracytoplasmic sperm injection (ICSI). Design: A retrospective analysis of 130 cyclesof ICSI performed for the treatment of male factor infertility. Setting: The Egyptian IVF-ET Center. Participants: One hundred thirty couples with the diagnosis of male factor infertility or with previous failed fertilization in conventional IVF or subzonal sperm injection. Intervention: Ovum pick-up and ICSI. Main Outcome Measure: Fertilization and pregnancy rates in relation to different semenparameters. Results: A total of 1,433oocytes were retrieved and 1,071 metaphaseII occytes

Citations from the literature /International

JOWMI

were injected. Normal fertilization occurred in 620 oocytes (58%).Embryo transfer was done for 128(98.5%)patients, and a total of 46 (35%) clinical pregnancies were achieved. There was no statistically signi!icant difference in the fertilization or pregnancy rates between patients who had previously failed fertilization in conventional IVF, patients with subfertile semen, patients with semenbetween 1 and 10 x 106/ml, and patients with semen < 1 x lob/ml. There was also no significant difference in the fertilization and pregnancy rates between patients with ~95% or >95% teratoxoospermia. Conclusion: In ICSI, the fertilization and pregnancy rates are not a&&d by different semen parameters as long as morphologically wellshaped live sperms could be used for the injection. -sperm~jcetioa:~-ofwrMe8iocoopkuwitbseverepllcfactoriofertilityisdependent primunyqmofenukuldmBtmakfactors Oehninger S.; Veeck L.; Lanxendorf S.; Maloney M.; Toner J.; Muasher S. USA FERTIL STERIL 19956415(977-981) Objective: To determine the efficacy and factors affecting outcome of intracytoplasmic sperm injection (RX) in patients with severemale factor infertility. Design: Prospectively designed clinical trial of patients selected to participate in the study basedupon the following inclusion criteria: previous total failed fertilization or unsuitable sperm parameters for conventional IVF. Setting: Tertiary care academic center. Patients: Ninety-two consecutive couples undergoing IVF therapy augmented with ICSI during April through December 1994 were studied. Main Outcome Measures:Fertilization and ongoing implantation and pregnancy rates (PRs). Results: A total of 1,163 preovulatory oocytes were manipulated, yielding a diploid fertilization rate of 60.9%; the oocyte damage rate was 13.2%.The transfer rate was 95% with 43.1% of cycles having excessembryos that were cryopreserved. Overall, the clinical and ongoing PRs per transfer were 31.9% and 26.8%, respectively. None of the sperm parameters of the original semen analysis correlated with ICSI outcome. Female age did not affect fertilization results but had a significant impact on PR (<34 years: 48.9%; 35 to 39 years: 22.90/o;240 years: 5.9% clinical PR per transfer). Conclusions: Intracytoplasmic sperm injection offers a new and powerful therapeutic option to treat couples with severemale factor infertility associatedwith a variety of sperm abnormalities. An adequate female age is a pivotal factor determining a successfuloutcome.

GYNECOLOGICAL

ONCOLOGY

Targeted eradkation of ovarian cancer mediatedby iotmxhh expressh of aati+bE2 &gIeeWn antIbedy Deshane J.; Cabrera G.; Grim J.E.; Siegal G.P.; Pike J.; Alvarez R.D.; Curie1 D.T. USA GYNECOL ONCOL 199559/l (8-14) Objective: Overexpression of the tyrosine kinase receptor

of Gynecology & Obstetrics 53 (19%) 305-316

311

erbB2 is important in the pathogenesis of a variety of neoplasms including ovarian cancer. As a strategy to selectively eradicate erbB2-overexpressing tumor cells, an anti-erbB2 single-chain itmntmoglobin (sFv) gene was constructed to direct expression of intracellular anti-erbB2 antibody. The purpose of this study is to establish the antitumorigenicity of this strategy in in vitro and in vivo models. Methods: An antierbB2 sFv construct containing an endoplasmic reticulum (ER)-directed leader sequencewas transiently expressedin the human ovarian carcinoma cell line SKOV3 using the adenovirus-polylysine vector. SKOV3 cells transfected with a non-ER form of an ant.i-erbB2 sFv construct or an irrelevant plasmid DNA served as controls. Antitumorigenicity, as measured by anchorage-independentgrowth in soft agar and subcutaneous(SC)tumor formation, was analyzed. The ability to achieve a biological effect with relevant sFv in murine orthotopic xenograft models and in primary human ovarian cancer cells was also evaluated. Results: The ER form of antierbB2 sFv was found to exert a marked antineoplastic effect on the SKOV3 cell line resulting in an arrest of anchorageindependent growth. A significant increasein SCtumor volume was noted in animals challenged with control constructs. In marked contrast, complete tumor eradication was noted at necropsy 80 days after sc transplantation in the group challenged with the ERdirected anti-erbB-2 sFv gene. Intraperitoneal treatment of malignant ascites in human tumor xenograft models with the ER form of the anti-erbb2 sFv gene resulted in profound down-regulation of cell surfaceerbB2 in retrieved ovarian cancer cells. The ERdirected anti-erbB2 sFv also elicited a significant cytotoxic effect in transfected primary ovarian cancer cells obtained from a patient with malignant ascites. Conclusion: The ability to selectively ‘knock out’ erbB-2 demonstratesthat this strategy can induce a significant antineoplastic effect in ovarian cancer cells overexpressing this growth factor receptor. In addition, the ability to accomplish selective abrogation of erbB-2 expression in animal treatment models and to transfect and eradicate primary ovarian cancer cellsjustifies further investigation of this novel strategy in ovarian cancer patients. Cembinatioa aati-ganetherapy targeting e-nryc and p53 In ovariallcanmcelIulles Jan&k M.F.; Sevin B.-U.; Nguyen H.N.; Averette H.E. USA GYNECOL ONCOL 199559/l (87-92) Gene therapy clinical trials targeting ~53 and other genesare underway in nongynecologic cancer systems. To explore the potential for antigene therapy in gynecologic oncology, we examined the in vitro effects of ohgonucleotides targeting c-myc and ~53 in the ovarian cancer cell lines CAOV3, SKOV-3, and BG-1. The ATP cell viability assay was used to measuregrowth effects after Cday treatments with 27-mer antisense phosphorothioate oligodeoxyribonucleotides (oligos) targeting the Puf/nm23 binding region of c-myc and promoter/ATG region of ~53. A random sequenceof the ~53 27-mer was usedas a control, and an untransformed tibroblast cell line