An effective strategy for rare microdissection testicular sperm involves utilizing a micro-cryopreservation technique...

The challenge with spermatozoa retrieval in non-obstructive azoospermia (NOA) patients is that testicular spermatozoa are frequently immotile and exceedingly scarce. The cryopreservation of these limited spermatozoa and the subsequent selection of a viable sperm from among immotile ones for intracytoplasmic sperm injection (ICSI) are critical steps. This study aimed to assess the clinical utility of a novel approach that combines a modified micro-cryopreservation technique with laser-assisted selection of immotile thawed sperm by microdissection testicular sperm extraction (micro-TESE). From January 2017 to December 2023, we performed a retrospective study comparing the outcomes of a micro-cryopreservation protocol combined with laser-assisted selection for micro-TESE-derived sperm from men with NOA (micro-TESE-ICSI group, n = 77) against those of fresh, motile TESA-derived sperm from men with obstructive azoospermia (TESA-ICSI group, n = 193). No significant differences were observed in the fertilization rate between the micro-TESE-ICSI and TESA-ICSI groups (P > 0.05), but good quality embryo rate and blastocyst rate of micro-TESE-ICSI were significantly reduced due to the worse sperm quality (P < 0.01). In fresh embryo transfer cycles, the micro-TESE-ICSI and TESA-ICSI groups showed no statistically significant differences in clinical pregnancy rate (38% vs 50%), implantation rate (29% vs 38%), miscarriage rate (14% vs 12%), or live birth rate (31% vs 38%) (P > 0.05 for all). Among patients who underwent a freeze-all embryos strategy, the first frozen-thawed embryo transfer cycle yielded comparable clinical pregnancy rates for the micro-TESE-ICSI and TESA-ICSI groups (70% vs 57%), with no statistically significant difference (P > 0.05). Similarly, no significant differences were observed in the implantation rate (61% vs 52%), miscarriage rate (4% vs 7%), or live birth rate (67% vs 48%) for the first frozen-thawed cycle (P > 0.05 for all). Furthermore, there was no statistically significant differences in mean gestational age, preterm delivery rate, mean birth weight at delivery, low birth weight and birth defects between the two groups in the first transfer cycle (fresh or frozen-thawed embryo transfer) (P > 0.05 in all cases). This study suggested that applying the modified micro-cryopreservation and laser selection protocol in micro-TESE-ICSI cycles may be a promising strategy for cases with scarce spermatozoa, as it resulted in pregnancy and neonatal outcomes similar to the use of fresh sperm from TESA.