B. Wu

Optimal use of fresh and frozen-thawed testicular sperm for intracytoplasmic sperm injection in azoospermic patients

Purpose: To optimize the use of fresh and frozen-thawed testicular biopsy specimens from patients with azoospermia.Methods: Fifty-one patients suffering from obstructive and non-obstructive azoospermia underwent testicular sperm extraction (TESE). The specimens were divided and used for either in vitro maturation or freezing for a future intracytoplasmic sperm injection (ICSI) cycle.Results: At initial testicular sperm extraction, very few motile spermatozoa were seen. After 24 h of in vitro maturation, sperm motility increased remarkably, with a maximum motility rate seen between 48 and 72 h of culture. Motile spermatozoa were observed up to 120 h in culture. In the 22 fresh ICSI cycles, a total of 294 oocytes were injected using motile sperm and 212 oocytes demonstrated normal 2PN formation (fertilization rate, 72.1%). In 36 frozen-thawed ICSI cycles, a total of 454 oocytes were injected and 302 oocytes became 2PN (66.5%). On day 3, high quality embryos were observed in 54.2% of fresh cycles and 54.1% of frozen cycles (P > 0.05). The clinical pregnancy rate did not show a significant difference between using fresh (59%) and frozen (55.5%) testicular biopsy sperm for ICSI (P > 0.05), but the embryo implantation rates did differ significantly between fresh (29.5%) and frozen-thawed (22.2%) cycles (P < 0.05). A total of 33 healthy babies have been born from 22 women, giving birth after 58 embryo transfer attempts (38%).Conclusion: The freezing and in vitro culturing of testicular biopsy tissue is a very reliable approach for the management of testicular biopsy specimens from azoospermic patients, and offers the possibility of several treatments of IVF/ICSI from a single sample.

Understanding reproducibility of human IVF traits to predict next IVF cycle outcome

PurposeEvaluating the failed IVF cycle often provides useful prognostic information. Before undergoing another attempt, patients experiencing an unsuccessful IVF cycle frequently request information about the probability of future success. Here, we introduced the concept of reproducibility and formulae to predict the next IVF cycle outcome.MethodsThe experimental design was based on the retrospective review of IVF cycle data from 2006 to 2013 in two different IVF centers and statistical analysis. The reproducibility coefficients (r) of IVF traits including number of oocytes retrieved, oocyte maturity, fertilization, embryo quality and pregnancy were estimated using the interclass correlation coefficient between the repeated IVF cycle measurements for the same patient by variance component analysis. The formulae were designed to predict next IVF cycle outcome.ResultsThe number of oocytes retrieved from patients and their fertilization rate had the highest reproducibility coefficients (r = 0.81 ~ 0.84), which indicated a very close correlation between the first retrieval cycle and subsequent IVF cycles. Oocyte maturity and number of top quality embryos had middle level reproducibility (r = 0.38 ~ 0.76) and pregnancy rate had a relative lower reproducibility (r = 0.23 ~ 0.27). Based on these parameters, the next outcome for these IVF traits might be accurately predicted by the designed formulae.ConclusionsThe introduction of the concept of reproducibility to our human IVF program allows us to predict future IVF cycle outcomes. The traits of oocyte numbers retrieved, oocyte maturity, fertilization, and top quality embryos had higher or middle reproducibility, which provides a basis for accurate prediction of future IVF outcomes. Based on this prediction, physicians may counsel their patients or change patient’s stimulation plans, and laboratory embryologists may improve their IVF techniques accordingly.

Improving ART pregnancy rates using culture with two kinds of media and two types of incubators

OBJECTIVE: Culture media and incubator systems have played a key role in IVF. Improved commercial culture media show little significant difference for embryo culture. However, we have observed individual patient’s embryos to have different responses for two media and two incubators. The objective of this study is to determine patient’s embryo response to culture medium and incubation. DESIGN: Prospective control study. MATERIALS AND METHODS: 1850 2PN zygotes from 220 patients were studied. In the same dish, patient’s embryos were divided. One sibling was placed in global medium and another in P1 medium. Each microdroplet contained a single embryo for culture. If patients had more than 4 embryos, they were cultured in two incubators with single CO2 gas or triple-gas. The cleavage rate and embryo quality on Day 2 and day 3 was recorded and the response of embryos to medium was determined based on embryo quality. Finally, pregnancy rates were determined in each group. RESULTS: The cleavage (global vs P1: 97.5% vs 98.7%), top quality embryos on Day 2 (74.1% vs 74.7%) and Day 3 (65.1% vs 60.4%) were not statistically different between media. However, different patient’s embryos had different responses to media. 40% (70/174) patient’s embryos grew very well in either global medium or P1 medium. 29% (50/174) patient’s embryos grew well only in P1with poor quality in global, while 20% (35/174) grew well in the global but poorly in the P1. 12% (21/ 174) did not grow well in both. The pregnant rate was 40% (10/25) in P1and 42.5% (9/21) in global (P>0.05). However, when two media were used simultaneously, the pregnant rate increased to 70.1% (122/ 174). Also, when two media were cultured in two incubators, it had a significant higher pregnant rate than in single incubator (73.2% vs. 60%, P<0.05). CONCLUSION: The favorable response of individual patient’s embryos to media and incubation suggest that in IVF clinical practice, using two media and two incubators for embryo culture could significantly improve IVF/ ICSI pregnant rates.