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The present study aimed to evaluate the clinical outcomes of magnetic-activated cell sorting (MACS) in sperm preparation for male subjects with a sperm DNA fragmentation index (DFI) ≥30%. A total of 86 patients who had undergone their first long-term long protocol were selected. The protocol involved in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI) cycles, and the patients were divided into the MACS or control groups. The MACS group included sperm samples analyzed with MACS that were combined with density gradient centrifugation (DGC) and the swim-up (SU) technique (n = 39), and the control group included sperm samples prepared using standard techniques (DGC and SU; n = 41). No differences were noted with regard to basic clinical characteristics, number of oocytes retrieved, normal fertilization rate, cleavage rate, or transplantable embryo rate between the two groups in IVF/ICSI. In addition, the clinical pregnancy and implantation rates of the first embryo transfer cycles indicated no significant differences between the two groups. However, there was a tendency to improve the live birth rate (LBR) of the first embryo transfer cycle (63.2% vs 53.9%) and the cumulative LBR (79.5% vs 70.7%) in the MACS group compared with the control group. Moreover, the number of transferred embryos (mean ± standard deviation [s.d.]: 1.7 ± 0.7 vs 2.3 ± 1.6) and the transfer number of each retrieved cycle (mean ± s.d.: 1.2 ± 0.5 vs 1.6 ± 0.8) were significantly lower in the MACS group than those in the control group. Thus, the selection of nonapoptotic spermatozoa by MACS for higher sperm DFI could improve assisted reproductive clinical outcomes. [ABSTRACT FROM AUTHOR]

Objective: To explore the relationship of the incidence of polynuclear zygotes with clinical pregnancy after short coincubation of gametes in in vitro fertilization (IVF)., Methods: We retrospectively analyzed 3 862 cases of short gamete coincubation IVF, which were divided into six groups according to the percentage of polynuclear zygotes: 0,1-10%, 11-20%, 21-30%, 31-50%, 41-50%, and > or = 51%. We compared the rates of clinical pregnancy, implantation and abortion among the six groups., Results: No statistically significant differences were found in the patients'age, dose of gonadotropin, peak E2, number of follicles at the hCG trigger, and number of oocytes retrieved among the six groups. The 1-10% group showed higher rates of pregnancy and implantation, while the > or = 51% group exhibited lower rates of pregnancy and implantation but a higher rate of abortion than the other groups, none with significant differences (P > 0.05)., Conclusion: The incidence of polynuclear zygotes after short coincubation of gametes in IVF cannot serve as a prognostic indicator of the outcome of clinical pregnancy.

Objective: To explore the effectiveness of ICSI in overcoming the high incidence of tripronucleates zygotes resulting from insemination in a previous IVF cycle., Methods: We retrospectively analyzed the matched-pair cycles in 37 patients with a > 35 % incidence of tripronucleate zygotes in an IVF cycle, with ICSI used in the subsequent cycle, evaluated the incidences of diploid (2PN) and triploid (3PN) zygotesand the number of normal embryos obtained, and compared the rates of clinical pregnancy and embryo implantation between the IVF and ICSI groups., Results: The mean age of the ICSI group was significantly older than that of the IVF group, while the ampules of gonadotropin and peak E2 showed no remarkable difference between the two. The numbers of follicles at hCG trigger, retrieved oocytes and mature oocytes were markedly lower in the former than in the latter. The percentage of 2PN was significantly higher while that of 3PN significantly lower after ICSI than after IVF (74.24% vs 34.42%; 11.57% vs 51.04%, P < 0.01), and more normal diploid embryos were obtained with ICSI (3.83 +/- 2.08 vs 2.52 +/- 1.71, P < 0.01). Four singletons were achieved in 31 IVF embryo transfer cycles, in comparison with 11 singletons and 3 twins in 36 ICSI embryo transfer cycles. The ICSI group showed significantly higher rates of clinical pregnancy and embryo implantation than the IVF group (38.89% vs 12.90%; 28.33% vs 7.41%, P<0.01)., Conclusion: For women with a high incidence o triploidy in a previous IVF cycle, ICSI can effectively increase the number of normal diploid zygotes.

Objective: To establish an effective pretreatment method for the vitrification of expanded mouse blastocysts by comparing 3 techniques for the artificial shrinkage of the blastocoelic cavity., Methods: The blastocoelic cavity was artificially shrunk by micro-needle aspiration, pipetting or laser drilling prior to the vitrification of the expanded blastocysts. The rates of survival and hatching achieved with the three techniques were compared with those of the non-shrinkage group., Results: The rates of survival were 72.9, 72.0 and 94.0%, and those of hatching were 64.6, 32.0 and 62.0% in the three shrinkage groups, obviously higher than in the nonshrinkage group (40.0 and 16.0%)., Conclusion: Artificial shrinkage of the blastocoelic cavity was an effective pretreatment technique for the vitrification of expanded mouse blastocysts, especially by micro-needle aspiration and laser drilling.

Objective: To report the derivation and characterization of a new human embryonic stem cell (hESC) line NJGLLhES1., Methods: From the inner cell mass of frozen-thawed human embryos and with ICR mouse embryonic fibroblasts as the feeder layer, we established a new human embryonic stem cell line, which was named NJGLLhES1. We detected the karyotype of the cell line, determined the expressions of alkaline phosphatase, the specific cell surface antigens SSEA-3, SSEA-4, TRA-1-60, TRA-1-81 and the marker gene Oct-4, and examined the formation of embryoids and teratomas., Results: NJGLLhES1 was maintained for over 1 year in vitro, with the morphological characteristics of hESC, a normal karyotype, positive expressions of alkaline phosphatase and specific cell marker genes, and the potential of forming embryoids and teratomas., Conclusion: A new human embryonic stem cell line NJGLLhES1 was successfully established, which remains karyotypically and phenotypically stable, undifferentiated and capable of self-renewal and pluripotential differentiation.

Objective: To investigate the influence of two different vitrification cryopreservation methods on the spindles of mouse M II oocytes., Methods: Three groups were included in the experiment, Group A, Group B and the control ( fresh oocytes). Mouse oocytes were vitrified by using cryoloop, with ethylene glycol( EG) in Group A and with EG + dimethyl sulphoxide ( DMSO) in Group B as cryoprotectants, and then the oocytes were placed directly into liquid nitrogen. Three hours after the frozen oocytes were thawed they were fixed, and the microtubule and chromosome were stained by indirect immunofluorescent method., Results: The survival rates of the oocytes after treated by the two vitrification cryopreservation methods had no difference ( 80. 3% vs 87. 5% , P > 0. 05) . The rate of the intact spindles in Group A was much lower than that of the control and Group B ( 15. 2% vs 78.7% , 15. 2% vs 77. 5% , P < 0. 05). But there was no difference between the latter two groups (78. 7% vs 77. 5% , P >0. 05). The oocytes with normal chromosome in Group A were much less than in the control and Group B (17.4% vs 76. 6% , 17. 4% vs 72. 5% , P <0. 05) , with no difference between the latter two groups(76. 6% vs 72. 5% , P >0. 05) ; The oocytes with abnormal chromosome were more in Group A than in the control and Group B (82. 6% vs 19. 1% , 82. 6% vs 27. 5% , P <0. 05) , with no difference between the latter two groups (19.1% vs 27.5% , P >0.05)., Conclusion: The changed vitrification cryopreservation method helps conserve the intact spindle configuration of mouse oocytes.