The analysis of sperm morphology can be used to evaluate sperm fertilizing ability and spontaneous conception status, and especially the overall analysis of the sperm head, neck and tail, along with the patient's living habits, occupation and clinical manifestations, may contribute to the primary diagnosis of the patients potentia generandi. It can also be employed to assess the effects of the treatment of semen samples. Although oocyte fertilization can be achieved by the technologies of intracytoplasmic sperm injection (ICSI), motile sperm organelle morphology examination (MSOME) and intracytoplasmic morphologically selected sperm injection (IMSI) regardless of sperm morphology and / or motility, which may somewhat weaken the clinical application of sperm morphology analysis, the standardized procedure and the practice of quality control for the analysis of sperm morphology can significantly improve the accuracy of its results and largely promote its clinical application. Therefore, it is of positive necessity as well as clinical application value to perform sperm morphology analysis in andrology laboratories, reproductive centers, sperm banks and the department of laboratory medicine.
Humans ; Male ; Semen Analysis ; Sperm Injections, Intracytoplasmic ; methods ; Sperm Motility

OBJECTIVETo analyze the clinical outcomes of intracytoplasmic sperm injection (ICSI) with spermatozoa from different sources.

METHODSWe retrospectively studied the rates of fertilization, clinical pregnancy, implantation, abortion, ectopic pregnancy and delivery in 682 patients treated by ICSI, who were divided according to the sperm sources into an ejaculated sperm group (ES, n = 598), a percutaneous epididymal sperm aspiration (PESA, n = 58) and a testicular sperm extraction (TSE, n = 26).

RESULTSThe fertilization rate was significantly lower in the TSE than in the ES and PESA groups (81.06% vs 87.95% and 87.82%, P < 0.05). But no statistically significant differences were observed among the ES, PESA and TSE groups in the rates of clinical pregnancy (39.46%, 48.28% and 34.62%), implantation (19.80%, 23.80% and 18.34%), abortion (13.13%, 17.86% and 11.11%), ectopic pregnancy (5.51%, 7.14% and 11.11%) and delivery (32.11%, 36.21% and 26.92%) (P > 0.05).

CONCLUSIONAlthough TSE-obtained sperm affect the rate of fertilization, those obtained by TSE and PESA do not obviously influence the outcome of clinical pregnancy.

Adult ; Female ; Humans ; Male ; Pregnancy ; Pregnancy Outcome ; Retrospective Studies ; Sperm Injections, Intracytoplasmic ; methods ; Sperm Retrieval ; Spermatozoa

OBJECTIVETo explore the effects of fertilization methods and sperm sources in intracytoplasmic sperm injection (ICSI) on the developmental capacity of surplus embryos.

METHODSWe analyzed the blastocyst formation of the surplus embryos from 2 135 patients, who were divided according to fertilization methods into an IVF (n=1803) and an ICSI group (n=332), the former again allocated to a normal fertilization (n=1642) and a rescue fertilization group (n=161), and the latter, according to sperm sources, to an ejaculated (n=248), an epididymal (n=70) and a testicular sperm group (n=14). The rates of blastocyst formation and good-quality blastocysts were compared between different fertilization methods and sperm sources.

RESULTSA total of 1884 blastocysts (28.87%) formed from 6525 surplus embryos of the patients after sequential culture, of which 974 (51.70%) were good-quality ones. The blastocyst formation rate of surplus embryos was significantly higher in the IVF (30.14%) than in the ICSI group (21.40%, P < 0.05), the rate of good-quality blastocysts was also higher in the former (52.44%) than in the latter (45.54%), but with no significant difference (P > 0.05). The rates of blastocyst formation and good-quality blastocysts were significantly higher in the normal (31.04% and 53.28%) than in the rescue fertilization IVF group (20.38% and 38.54%, P < 0.05), and in the testicular sperm ICSI group (30.23% and 53.85%) than in either the epididymal (18.36% and 42.11%) or the ejaculated sperm ICSI group (21.76% and 45.70%) (P < 0.05).

CONCLUSIONThe development potential of surplus embryos was higher in IVF than in ICSI, in the normal than in the rescue fertilization IVF group, and in the testicular than in the epididymal and ejaculated sperm ICSI groups.

Blastocyst ; Embryonic Development ; Female ; Fertilization in Vitro ; methods ; Humans ; Male ; Oocytes ; Sperm Capacitation ; Sperm Injections, Intracytoplasmic ; methods ; Sperm Motility

This paper describes the use of piezo-driven micropipette for intracytoplasmic sperm injection of mice eggs. The head of fresh spermatozoa from KM (Kunming) fertile mice was individually injected into mature oocytes of hybrid mice B6D2F1. Approximately eighty three percent of sperm-injected oocytes survived, and 84.0% of them fertilized normally (extrusion of the second polar body and formation of male and female pronuclei). The eggs fertilized by sperm injection could develop in vitro to 2-cell (98% vs 94.7%), 4-cell (89.5% vs 92.1%) stages, no significantly (P > 0.05) different from embryos fertilized in vivo but there were significantly (P < 0.01) few morulae (63.8% vs 84.2%) and blastocysts (25.7% vs 68.4%) developed in vitro after further culture in vitro in the group of ICSI. When 120 embryos at the pronuclear stage were transferred to seven pseudopregnant KM female, 23.3% of the embryos (0 - 50%, depending on the host) reached the full term. Except for three that were cannibalized soon after birth, all of the young (25 pups) developed into normal and fertile adult. Here we report the first birth of mouse offspring following ICSI in China. These studies may increase understanding of the fertilization process and of how ICSI works.
Animals ; Embryo Transfer ; Female ; Fertilization in Vitro ; methods ; Male ; Mice ; Oocytes ; physiology ; Pregnancy ; Sperm Injections, Intracytoplasmic ; methods

Sperm selection plays an important role in assisted reproductive technology. In recent years, sperm evaluation is not limited to the assessment of sperm motility and morphology, but involves more other sperm characteristics such as sperm ultrastructure, DNA integrity, apoptosis and membrane. Assessment based on these characteristics is becoming the aim of sperm selection. This article gives an overview on several newly developed techniques for sperm selection according to different technical principles, such as electrophoretic separation, zeta potential, HA binding, Annexin V binding, intracytoplasmic morphologically selected sperm injection (IMSI) and microfluidic sperm sorter, which have all been applied to IVF or ICSI with the exception of microfluidic sperm sorter. It also introduces the advantages, disadvantages and application effects of these techniques.
Cell Separation ; Fertilization in Vitro ; methods ; Humans ; Male ; Reproductive Techniques, Assisted ; Semen Analysis ; Sperm Injections, Intracytoplasmic ; methods

OBJECTIVETo determine an optimal insemination technique for patients suspected of high risk of fertilization failure and undergoing assisted reproduction treatment.

METHODSNinety-nine couples were treated by conventional in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI) in one cycle (half-ICSI) by dividing the sibling oocytes in halves. The clinical and laboratory data were analyzed, and the rates of fertilization, cleavage, good embryos and clinical pregnancy were compared between different fertilization methods.

RESULTSIn the half-ICSI group, the fertilization rate of ICSI (80.5%) was significantly higher than that of IVF (42.9%) (P < 0.01), and so were the rates of complete fertilization failure (21.2%) and low fertilization (16.2%) of IVF than those of ICSI (0 and 3.0%). No significant differences were observed in the rates of cleavage and good-quality embryos between the two groups (P > 0.05).

CONCLUSIONICSI can help to avoid complete fertilization failure, achieve more high quality embryos for transfer and improve the rate of pregnancy for patients with high risk of fertilization failure.

Female ; Fertilization in Vitro ; methods ; Humans ; Male ; Oocytes ; cytology ; Pregnancy ; Pregnancy Outcome ; Sperm Injections, Intracytoplasmic ; methods

Total or near-total fertilization failure after intracytoplasmic sperm injection (ICSI) is a rare event, but it occurs repeatedly because of sperm defects in activating oocyte. The case presents a successful pregnancy and live birth after calcium ionophore A23187 (A23187) activation on one-day-old unfertilized oocytes in a patient whose husband suffered oligoasthenoteratozoospermia, and who had experienced repeated near-total fertilization failure after ICSI. In the second ICSI cycle, only one oocyte was fertilized while nine were unfertilized. Oocyte activation with A23187 were performed on the one-day-old unfertilized oocytes after ICSI and resulted in fertilization and embryo transfer. A clinical pregnancy was achieved and a healthy baby was born. To our knowledge, this is the first reported case of a healthy birth after oocyte activation on the one-day-old unfertilized oocyte. This indicates that "rescue oocyte activation" on one-day-old unfertilized oocytes after ICSI may be helpful for preventing total or near-total fertilization failure after ICSI.
Adult ; Female ; Fertilization in Vitro ; methods ; Humans ; Live Birth ; Male ; Oocytes ; cytology ; Pregnancy ; Sperm Injections, Intracytoplasmic ; methods

After freeze-drying, the ultrastructure of boar sperms was observed by optical and electron microscopy. The in vitro development ability of the sperm was also examined with intracytoplasmic sperm injection (ICSI). The rate of male pronuclear formation was (68.52%), for cleavage (59.17%) and for blastocyst formation (19.16%) of the trehalose group (0.2 mol/L), significantly higher than those of the 50 mmol/L EDTA group (64.59%, 56.26% and 15.62%) and the control group (35.36%, 52.33% and 8.60%) (P < 0.05). After storage for 60, 120 and 180 d at 4 degrees C, no significant difference in the in vitro development was observed (P > 0.05). The male pronuclear, cleavage and blastocyst formation after ICSI with freeze-dried spermatozoa incubated for 1 h was superior than those incubated for 2 h (P < 0.05). No significant differences in the structures after stored at 4 degrees C or -20 degrees C (P > 0.05) were observed between the trehalose group and EDTA group. The percent of B grade freeze-dried boar spermatozoa in the trehalose group was higher than that of the EDTA group (P < 0.05). Based on the ultrastructure observation, main cryogenic damage in freeze-dried boar spermatozoa was swelling, damage or rupture in the sperm acrosome, neck and tail.
Animals ; Freeze Drying ; Male ; Semen Preservation ; methods ; veterinary ; Sperm Injections, Intracytoplasmic ; veterinary ; Spermatozoa ; Swine ; Trehalose ; pharmacology

OBJECTIVETo assess whether intracytoplasmic morphologically selected sperm injection (IMSI) of testicular sperm improves the clinical outcome in patients with azoospermia.

METHODSWe performed conventional intracytoplasmic sperm injection (ICSI) for 66 patients diagnosed with azoospermia and IMSI for another 39 using testicular sperm selected at high magnification ( x 6000), and comparatively analyzed the clinical outcomes of the two techniques.

RESULTSThere were no statistically significant differences between conventional ICSI and IMSI in the rates of pregnancy (51.52% vs. 56.41%) and implantation (30.67% vs. 35.29%), although the rate of early abortion was lower in the IMSI than in the ICSI group (4.50% vs. 11.76%).

CONCLUSIONIMSI of testicular sperm may effect a lower rate of early abortion than conventional ICSI in patients with azoospermia.

Adult ; Azoospermia ; therapy ; Female ; Humans ; Male ; Pregnancy ; Pregnancy Rate ; Sperm Injections, Intracytoplasmic ; methods ; Treatment Outcome

In our previous study, normal and fertile mice were successful produced from oocytes following intracytoplasmic sperm injection (ICSI). In the present study, the possibility of producing transgenic embryos and offspring with this procedure was evaluated. After freezing-thawed once using HEPES-CZB medium without cryoprotectants, the cauda sperm from KM fertile male were exposed to the circular or linear pEGFP-N1 DNA for 1 min and then co-injected into metaphase II oocytes of B6D2F1 strain. When the zygotes with two pronuclei were cultured in CZB medium to day 3.5, 39.1% (9/23) of them, derived from oocytes co-injected with sperm head and pEGFP-N1 plasmid DNA, were expressed GFP protein. After transfer of the ICSI embryos with two pronuclei from co-injection of sperm head and foreign DNA, seven recipients delivered 30 pups (23.8%, 30/126). Southern blot results revealed that three of sixteen offspring integrated with GFP and neomycin genes together (18.8 %). Interestingly, all of them were produced from oocytes co-injected sperm head and linear DNA (33.3%, 3/9), while none of seven ICSI offspring integrated either GFP or neomycin gene in the group of co-injection of sperm head and circular plasmid DNA. These results indicated that the high efficiency of transgenic mouse could be produced by ICSI. It may be shown that linear DNA is more easily to integrate into host genome than circular DNA when ICSI was used to produce transgenic animals.
Animals ; Male ; Mice ; Mice, Inbred C57BL ; Mice, Inbred DBA ; Mice, Transgenic ; genetics ; Sperm Injections, Intracytoplasmic ; methods