常規使用PVP減低精蟲活動力，研究顯示不能暴露時間超過15分鐘，超過會對於精蟲DNA染色體有不量影響

Prolonged exposure of human spermatozoa in polyvinylpyrrolidone has detrimental effects on sperm biological characteristics

Polyvinylpyrrolidone (PVP) has been utilized in intracytoplasmic sperm injection (ICSI) for immobilization and manipulation of spermatozoa. This study aims to determine the suitable time that sperm cells could be safely exposed to PVP during ICSI procedure. Twenty-five normal semen samples were prepared using the swim-up method and then were exposed to 10% PVP at different time intervals (15, 30 and 60 min). The effect of PVP on sperm parameters (viability and morphology), DNA fragmentation index (sperm chromatin dispersion test), chromatin quality (aniline blue, toluidine blue and chromomycin A3 staining), acrosome reaction, mitochondrial membrane potential and sperm ultrastructure was assessed at different time intervals. Our results showed that prolonged sperm exposure in PVP for 15, 30 and 60 min significantly affects viability and morphology with a concomitant increase in DNA fragmentation and abnormal chromatin structure, while the percentage of acrosome-reacted spermatozoa was additionally increased. In addition, the spermatozoa with high mitochondrial membrane potential were significantly decreased compared to unexposed spermatozoa to PVP. In conclusion, the detrimental effects of PVP were increased significantly following sperm exposure in PVP after 15 min. Therefore, the sperm exposure to PVP should be limited to less than 15 min during ICSI procedure.

 PVP使用於精蟲顯微注射之精蟲遲滯

傳統PVP濃度為7%，研究顯示較低濃度之PVP (5%)可能有助於提高胚胎

Effects of different polyvinylpyrrolidone concentrations on intracytoplasmic sperm injection

Published online by Cambridge University Press:  14 January 2020

Summary

To explore whether different polyvinylpyrrolidone (PVP) concentrations affect the results of intracytoplasmic sperm injection (ICSI), a prospective study was conducted for 194 couples undergoing 210 ICSI therapy cycles. These cycles were divided into three groups (10, 7 and 5% groups) using the corresponding concentration of PVP for sperm immobilization. The main outcome measures were analyzed. Results indicated that, with a decrease in PVP concentrations, all of the main outcome measures increased. In particular, the high-quality cleavage embryo rate in the 7% group was significantly lower than in the 5% group (P < 0.01), and the cleavage, high-quality cleavage embryo, and high-quality blastocyst rates in the 5% group were significantly higher than those in the 10% group (all P < 0.001). For high-/intermediate-quality semen, all of the main outcome measures were significantly increased with 5% PVP. For the poor-quality semen, only the high-quality cleavage embryo and high-quality blastocyst rates were significantly higher in the 5% group. Therefore, lowering PVP concentrations greatly promoted the development of embryos in ICSI cycles, with an optimal concentration of 5% for ICSI.

是否D3更新培養液並無臨床差異

one step   vs two step 囊胚率: 68.3 vs. 66.8%)

Fertil Steril  2016 Mar;105(3):707-713.

 doi: 10.1016/j.fertnstert.2015.11.038. Epub 2015 Dec 12.

Blastocyst development in single medium with or without renewal on day 3: a prospective cohort study on sibling donor oocytes in a time-lapse incubator

Objective: To evaluate the efficiency of using a continuous (one-step) protocol with a single medium for the culture of human embryos in a time-lapse incubator (TLI).

Patient(s): Embryos from 59 patients.

Intervention(s): Culture in a TLI in a single medium with or without renewal of the medium on day-3.

Main outcome measure(s): Embryo morphology and morphokinetic parameters, clinical pregnancy, take-home baby rate, and perinatal outcomes.

Result(s): The blastocyst rates (68.3 vs. 66.8%) and the proportion of good-quality blastocysts (transferred plus frozen) obtained with the two-step (80.0%) protocol were statistically significantly similar to those obtained in the one-step protocol (72.2%). Similarly, morphokinetic events from early cleavage until late blastocyst stages were statistically significantly equivalent between both groups. No differences were found either in clinical pregnancy rates when comparing pure transfers performed with embryos selected from the two-step (75.0%), one-step (70.0%, respectively), and mixed (57.1%) groups. A total of 55 out of 91 embryos transferred implanted successfully (60.4%), resulting in a total of 37 newborns with a comparable birth weight mean among groups.

Conclusion(s): Our findings support the idea that in a TLI with a controlled air purification system, human embryos can be successfully cultured continuously from day 0 onward in single medium with no need to renew it on day-3. This strategy does not affect embryo morphokinetics or development to term and offers more stable culture conditions for embryos as well as practical advantages and reduced costs for the IVF laboratory.