Assisted fertilization by subzonal insemination and intracytoplasmic sperm injection

1994 ◽  
Vol 6 (1) ◽  
pp. 85 ◽  
Author(s):  
Steirteghem A Van ◽  
J Liu ◽  
H Joris ◽  
Z Nagy ◽  
C Staessen ◽  
...  
Keyword(s):  
Intracytoplasmic Sperm Injection ◽  
Congenital Malformations ◽  
Subsequent Development ◽  
Fertilization Rate ◽  
Subzonal Insemination ◽  
Failed Fertilization ◽  
Assisted Fertilization ◽  
Normal Fertilization

The results of 600 consecutive treatment cycles of subzonal insemination (SUZI) and intracytoplasmic sperm injection (ICSI) are described in couples with failed fertilization after standard IVF or insufficient spermatozoa in the ejaculate for IVF. More oocytes were damaged by ICSI (16.3%) than by SUZI (8.5%) and the normal fertilization rate was substantially higher after ICSI (49.1% v. 16.6%). Subsequent development of two-pronuclear oocytes in vitro was 80% after SUZI and 73.9% after ICSI. Significantly more triple embryo replacements were carried out after ICSI than after SUZI. Embryo transfers were possible in 421 of the 600 cycles. There were 63 pregnancies after ICSI (215 transfers) and 23 after SUZI (156 transfers); 10 additional pregnancies were achieved after 50 transfers of a mixture of SUZI and ICSI embryos. The results of fetal karyotypes and follow-up of the children do not indicate an increase in congenital malformations.

Clinical results from intracytoplasmic sperm injection at monash IVF

1995 ◽  
Vol 7 (2) ◽  
pp. 247 ◽  
Author(s):  
RI McLachlan ◽  
G Fuscaldo ◽  
H Rho ◽  
C Poulos ◽  
J Dalrymple ◽  
...  
Keyword(s):  
Intracytoplasmic Sperm Injection ◽  
Oocyte Retrieval ◽  
Clinical Results ◽  
Fertilization Rate ◽  
Vitro Fertilization ◽  
Failed Fertilization ◽  
Patient Groups ◽  
Sperm Antibodies ◽  
The Impact

The impact of a modification of the intracytoplasmic sperm injection (ICSI) technique on fertilization and pregnancy rates was examined in a retrospective analysis of 171 consecutive ICSI treatment cycles (156 patients). Patients were selected for ICSI on the basis of severe oligoasthenozoospermia (65 patients) or following conventional in vitro fertilization (IVF) with failed or poor fertilization (70 patients). Seven patients in which epididymal or testicular sperm was used, 10 patients with sperm antibodies and 4 patients with retrograde ejaculation or who required electro-ejaculation were also treated with ICSI. In the first 105 cycles (102 patients), single sperm, rendered immotile, were injected into the ooplasm of 979 metaphase II (M II) oocytes using an established technique (Method 1). In the following 66 cycles (513 M II oocytes injected), the ICSI procedure was modified by increased aspiration of the oolemma to ensure the intracytoplasmic deposition of sperm (Method 2). The patient groups did not differ between the two injection procedures. The normal (two pronuclear) fertilization rate increased significantly (P < 0.001) from 34.3% with Method 1 to 73.1% with Method 2, with no difference in the oocyte degeneration rate (4.3% v. 4.5% respectively). The incidence of failed fertilization was significantly (P < 0.01) reduced from 17.1% (18 cycles) to 1.6% (1 cycle) with the change in technique. As a consequence of the increased fertilization rates with Method 2, more embryos were available for assessment and transfer, and a pregnancy rate per oocyte retrieval of 21.2% was obtained for Method 2. Fertilization, embryo transfer and pregnancies were obtained in all patient groups treated with ICSI.(ABSTRACT TRUNCATED AT 250 WORDS)

Intracytoplasmic sperm injection — ICSI

1994 ◽  
Vol 3 (3) ◽  
pp. 199-207 ◽  
Author(s):  
AC Van Steirteghem ◽  
P Nagy ◽  
J Liu ◽  
H Joris ◽  
J Smitz ◽  
...  
Keyword(s):  
Fertilization Rate ◽  
Male Factor Infertility ◽  
Male Factor ◽  
The Past ◽  
Partial Zona Dissection ◽  
Tubal Disease ◽  
Sizeable Number ◽  
Assisted Fertilization ◽  
Normal Fertilization

For more than a decade in vitro fertilization (IVF) has been successful in the treatment of couples with long-standing infertility due to various aetiologies such as tubal disease, male-factor infertility, unexplained infertility and endometriosis. The usual fertilization rate in IVF for nonmale infertility cases is 60–70% of the inseminated cumulus-oocyte complexes and in andrological infertility it is only 20–30%. The lower the number of normally fertilized oocytes, the less chance there is of available embryos, so that patients may have no embryos to transfer. It has been the experience of all centres for reproductive medicine, including our own, that a certain number of couples with male-factor infertility cannot be helped by standard IVF treatment. After insemination with progressively motile spermatozoa the number of two-pronuclear oocytes was either zero or less than 5%. Furthermore, a sizeable number of couples cannot be accepted for IVF if the number of progressively motile spermatozoa in the ejaculate is below a certain threshold number such as 500 000. In the past five years, assisted fertilization procedures have been developed to circumvent the barriers that prevent sperm access to the ooplasma, namely the zona pellucida and the ooplasmic membrane. Pregnancies and births have been reported after partial zona dissection (PZD) and subzonal insemination (SUZI). The success rate of PZD and SUZI has remained moderate: the normal fertilization rate (two-pronuclear oocytes) has never exceeded 20–25% of the micromanipulated oocytes; only two-thirds of the patients have had embryo transfers of, usually, a low number of embryos, resulting in a reduced pregnancy and take-home baby rate.

Effects of oocyte age, cumulus cells and injection methods on in vitro development of intracytoplasmic sperm injection rabbit embryos

Zygote ◽  
2004 ◽  
Vol 12 (1) ◽  
pp. 75-80 ◽  
Author(s):  
Yue-Liang Zheng ◽  
Man-Xi Jiang ◽  
Yan-Ling Zhang ◽  
Qing-Yuan Sun ◽  
Da-Yuan Chen
Keyword(s):  
Intracytoplasmic Sperm Injection ◽  
Polar Body ◽  
Cumulus Cells ◽  
Fertilization Rate ◽  
First Polar Body ◽  
Injection Methods ◽  
Blastocyst Rate ◽  
Repeated Aspiration ◽  
Rabbit Embryos

This study assessed the effects of oocyte age, cumulus cells and injection methods on in vitro development of intracytoplasmic sperm injection (ICSI) rabbit embryos. Oocytes were recovered from female rabbits superovulated with PMSG and hCG, and epididymal sperm were collected from a fertile male rabbit. The oocyte was positioned with the first polar body at 12 o'clock position, and a microinjection needle containing a sperm was inserted into the oocyte at 3 o'clock. Oolemma breakage was achieved by aspirating ooplasm, and the aspirated ooplasm and sperm were re-injected into the oocyte. The injected oocytes were cultured in M199 medium containing 10% fetal calf serum at 38 °C with 5% CO2 in air. The results showed that oocytes injected at 1 h post-collection produced a higher (p<0.05) fertilization rate than those injected at 4 or 7 h post-collection. Blastocyst rate in the 1 h group was higher (p<0.05) than in the 7 h group. Denuded oocytes (group A) and oocytes with cumulus cells (group B) were injected, respectively. Rates of fertilization and development of ICSI embryos were not significantly different (p<0.05) between the two groups. Four ICSI methods were applied in this experiment. In methods 1 and 2, the needle tip was pushed across half the diameter of the oocyte, and oolemma breakage was achieved by either a single aspiration (method 1) or repeated aspiration and expulsion (method 2) of ooplasm. In methods 3 and 4, the needle tip was pushed to the oocyte periphery opposite the puncture site, and oolemma breakage was achieved by either a single aspiration (method 3) or repeated aspiration and expulsion (method 4) of ooplasm. Fertilization rate in method 2 was significantly higher (p<0.05) than in methods 1 and 3. Blastocyst rates were not significantly different (p<0.05) among methods 1, 3 and 4, but method 2 produced a higher (p<0.05) blastocyst rate than method 3.

Experience with assisted fertilization in severe male factor infertility and unexplained failed fertilization in vitro

1994 ◽  
Vol 9 (4) ◽  
pp. 680-683 ◽  
Author(s):  
C.J. Redgment ◽  
D. Yang ◽  
M. Tsirigotis ◽  
N. Yazdani ◽  
T. AI Shawaf ◽  
...  
Keyword(s):  
Fertilization In Vitro ◽  
Male Factor Infertility ◽  
Male Factor ◽  
Failed Fertilization ◽  
Assisted Fertilization

Development and implementation of intracytoplasmic sperm injection (ICSI)

1995 ◽  
Vol 7 (2) ◽  
pp. 211 ◽  
Author(s):  
GD Palermo ◽  
J Cohen ◽  
M Alikani ◽  
A Adler ◽  
Z Rosenwaks
Keyword(s):  
Intracytoplasmic Sperm Injection ◽  
Human Sperm ◽  
Fertilization Rate ◽  
Semen Parameters ◽  
Ongoing Pregnancy Rate ◽  
Vitro Fertilization ◽  
Human Oocytes ◽  
Intracytoplasmic Injection ◽  
Previously Treated

The purpose of this paper is to elucidate the experimental steps that led to the development of intracytoplasmic sperm injection (ICSI) and its application in the human. ICSI has become the most successful micromanipulation procedure for treating male infertility. A total of 355 in vitro fertilization (IVF) cycles utilizing ICSI are described; 180 couples were previously treated in 509 IVF cycles but achieved no fertilization and 175 couples could not be treated by IVF because of extremely poor semen parameters. Of the 3063 metaphase II (M II) oocytes retrieved, 2970 were injected with a survival rate of 93.6%, yielding 1917 bipronuclear zygotes (64.5%). In 148 patients, a foetal heart was evidenced by ultrasound; 11 of these patients miscarried between 7 and 13 weeks of gestation. The ongoing pregnancy rate was 38.6% (137/355) per retrieval and 40.5% (137/338) per embryo replacement. At the time of writing, there were 22 deliveries and one therapeutic abortion for a trisomy 21 chromosomal abnormality. In addition, 66 singleton, 37 twin, 10 triplet and 1 quadruplet pregnancies were ongoing. The concentration of motile spermatozoa in the ejaculate only slightly influenced the fertilization rate (P < 0.001) and the pregnancy outcome (P < 0.01). A preliminary injection procedure utilizing intracytoplasmic injection of isolated sperm heads was performed in 35 M II human oocytes with resultant fertilization and cleavage rates of 74% and 73% respectively. Skills in ICSI were acquired by injecting hamster and unfertilized human oocytes with human sperm. ICSI can be used to successfully treat couples who have failed IVF or who have too few spermatozoa for conventional in vitro insemination.(ABSTRACT TRUNCATED AT 250 WORDS)

320 EFFECTS OF DIFFERENT ACTIVATION REGIMENS ON PRONUCLEAR FORMATION AND PRE-IMPLANTATION DEVELOPMENTAL COMPETENCE OF IN VITRO-MATURED BOVINE OOCYTES AFTER INTRACYTOPLASMIC SPERM INJECTION

2015 ◽  
Vol 27 (1) ◽  
pp. 249
Author(s):  
M. E. Arias ◽  
R. Sanchez ◽  
R. Felmer
Keyword(s):  
Intracytoplasmic Sperm Injection ◽  
Standard Procedure ◽  
Chemical Activation ◽  
Developmental Rate ◽  
Fertilization Rate ◽  
Developmental Competence ◽  
Assisted Reproductive Technique ◽  
Pronuclear Formation ◽  
Bovine Oocytes

Intracytoplasmic sperm injection (ICSI) is an assisted reproductive technique that has been used with considerable success in humans; however, in the bovine species the efficiency of this technique is far from optimal. The objective of the present study was to evaluate the effect of 4 chemical activation treatments, 6-dimethylaminopurine (DMAP), cycloheximide (CHX), anisomycin (ANY), and ethanol (EtOH) on the pronuclear formation and embryo development of bovine embryos generated by ICSI. Cumulus-oocyte complexes were aspirated from abattoir ovaries, selected, and matured in 400-µL drops of standard TCM-199 maturation medium for 22 h at 38.5°C and 5% CO2. The ICSI was performed by a standard procedure. Injected oocytes were randomly distributed and activated by 5 µM ionomycin for 5 min (Io) followed by i) 5 µg mL–1 CHX for 5 h (Io/CHX), ii) 3 h window followed by a second Io treatment plus 1.9 mM DMAP for 4 h (2Io/DMAP), iii) 1 µg mL–1 ANY for 5 h (Io/ANY), and iv) 3 h window followed by 7% ethanol (Io/EtoH). Embryos were cultured in 50-µL drops of KSOM medium under mineral oil at 38.5°C and 5% CO2, 5% O2, and 90% N2. Cleavage was recorded at 72 h and blastocyst rate at 192 h. Pronuclear formation analysis was carried out at 18 hpa with Hoechst staining. An oocyte was considered fertilized when 2 polar bodies and 1 female and 1 male pronucleus (or a decondensed sperm head) could be observed. The data were transformed to arcsine, analysed by ANOVA, and means were compared using Tukey's test with Statgraphics Plus 2 Software. Results with a total of 431 injected oocytes (114, 104, 101, and 112 for DMAP, CHX, ANY, and EtOH, respectively) showed differences in cleavage (P < 0.01) in DMAP, CHX, and ANY treatments (86, 72, and 78%, respectively), relative to EtOH (12%). Similarly, the rate of blastocysts/injected oocyte at 192 h was higher with DMAP, CHX, and ANY (41, 20, and 32%, respectively), relative to EtOH (4%). Sham-injected oocytes showed cleavage and blastocyst rates of 67, 43, 68, and 12% and 32, 11, 19, and 5%, for DMAP, CHX, ANY, and EtOH, respectively. Despite the higher developmental rate observed with DMAP, pronuclear formation assessment revealed that fertilization rate was higher in CHX (87%) and ANY (75%) treatments relative to DMAP (35%). In conclusion, the results of the present study show that activation of bovine oocytes after ICSI is more efficient with DMAP and ANY, compared with CHX and EtOH.Provision of ovaries by our local slaughterhouse (Frigorifico Temuco, Chile) and funding support from FONDECYT 1120241 CONICYT, Chile, are gratefully acknowledged.

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