Effect of Early Cumulus Cell Removal on the Fertilization and Clinical Outcome in Human <i>in Vitro</i>Fertilization

The short-term insemination technique has been widely applied, although it is not clear whether the mechanical desorption of cumulus cells can affect normal fertilization and birth outcomes. Therefore, the present study aimed to evaluate whether early cumulus cell removal could affect in vitro fertilization outcomes. Based on a history of infertility and semen analysis, 61 patients were identified, and the cumulus cells were mechanically removed at either6 h post-insemination (Group A, 387 mature oocytes) or 18 - 20 h post-insemination as traditional in vitro fertilization (Group B, 424 mature oocytes). There were no significant differences in the rates of fertilization, available embryos, high quality embryos, birth, premature birth, pregnancy, or implantation when we compared the two groups. However, Group A exhibited a significantly higher polyspermy rate, and significantly lower rates of blastocyst formation and high-quality blastocyst formation, compared with Group B. Therefore, the results of present study indicate that early cumulus cell removal (at 6 h post-insemination) may negatively affect the rates of polyspermy, blastocyst formation, and high-quality blastocyst formation.

Presence of Meiotic Spindles Indicates Early Cleavage of Embryos

Objective To assess whether the detection of the meiotic spindle could anticipate the appearance of early cleavage. Methods Oocytes were obtained from stimulated ovaries of consenting patients undergoing oocytes retrieval for ICSI. Spindles were imaged with the Polscope. After ICSI, oocytes with or without spindles were cultured for examination of early cleavage and embryo development. A total of 328 oocytes from 50 cycles were examined with the Polscope and inseminated by ICSI. Results Spindles were imaged in 81.7% of oocytes. After ICSI, more oocytes with spindles （78.4%） fertilized normally than oocytes without spindles （53.3%） （P〈0. 001）. At 25-27 h post ICSI, more fertilized oocytes developed from oocytes with spindles （81.9%） were detected early cleavage than those from oocytes without spindles （28.1%） （P〈0.001）. Significantly more embryos with early cleavage （82.2%） developed to high quality embryos at d 3 compared with the embryos without early cleavage （48.3%） （P=0.001）. The value of rs related to the relationship between spindles and early cleavage was 0.420 （P〈0.001）. Conclusion The existing of the early cleavage may have a predictive value on the opportunity of high quality embryos and the existing of the spindle may have a predictive value in the appearance of early cleavage.

Effects of short gamete co-incubation in human in vitro fertilization

Traditionally oocytes have been exposed to sperm overnight for 16-20 h.This long period of co-incubation, however,has been shown to create problems with high levels of reactive oxygen species(ROS),which may affect embryo viability and cause hardening of the zona pellucida.Recently,several positive and negative effects of reducing the co-incubation time to 90-120 min were reported.The objective of this review was to discuss whether short exposure of oocytes to sperm in vitro might affect the clinical results and several problems arose in short gamete co-incubation.

Novel DPY19L2 variants in globozoospermic patients and the overcoming this male infertility

Globozoospermia has been reported to be a rare but severe causation of male infertility,which results from the failure of acrosome biogenesis and sperm head shaping.Variants of dpy-19-like 2(DPY19L2)are highly related to globozoospermia,but related investigations have been mainly performed in patients from Western countries.Here,we performed a screening of DPY19L2 variants in a cohort of Chinese globozoospermic patients and found that five of nine patients carried DPY19L2 deletions and the other four patients contained novel DPY19L2 point mutations,as revealed by whole-exome sequencing.Patient 3(P3)contained a heterozygous variant(c.2126+5G>A),P6 contained a homozygous nonsense mutation(c.1720C>T,p.Arg574*),P8 contained compound heterozygous variants(c.H82-1184delATCf p.Leu394_Ser395deIinsPhe;c.368A>T,p.Hisl23Arg),and P9 contained a heterozygous variant(c.H82-1184delATCTTI frameshift).We also reported intracytoplasmic sperm injection(ICSI)outcomes in the related patients,finding that ICSI followed by assisted oocyte activation(AOA)with calcium ionophore achieved high rates of live births.In summary,the infertility of these patients results from DPY19L2 dysfunction and can be treated by ICSI together with AOA.

Effects of Different Serum on in vitro Cultured Human Granulosa Cells

Objective To evaluate the effects of different sera on the growth of human granulosa cells （GCs） cultured in vitro. Methods GCs were obtained from women who underwent follicular aspirates during in vitro fertilization （IVF） program. Five groups were divided according to media supple- mented with different types of sera. Group A, 10% heat-inactivated fetal bovine serum （FBS）; group B, 10% heat-inactivated newborn bovine serum; group C, 10% non heat-inactivated FBS; group D, 10% human serum; group E, bovine serum albumin. Morphological characteristics and viability of GCs measured by trypan blue exclusion assay were evaluated after 24 h of incubation.Results GCs cultured in group A and group B showed multiform morphology compared with other groups. GCs cultured in group D and group E were present with cytoplasmic atrophy and less pseudopodium. Moreover, group A and group B showed a similar level in the viability of GCs （P〉0.05）, which displayed no difference between group D and group E as well （P〉0.05）. Group C had a lower level of viability than group A （P〈0.05） but a higher level than group D （P〈0.01）. Conclusion Heat-inactivated sera can improve the growth of GCs. Different types of sera would have different effects on the growth of GCs cultured in vitro. The pre-culture with different types of sera should be performed to get better efficacy.