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A Novel Three-Step Protocol for Laboratory Processing of Testicular Tissue in Non-Obstructive Azoospermia Patients
Xuefeng Luo1,2, Chen Luo1,2, Chengchao Du1,2
1Department of Reproductive Andrology & Sichuan Human Sperm Bank, West China Second University Hospital, Sichuan University, Chengdu, China.
Andrology
|December 31, 2025
Summary
Establishing a three-step laboratory process for testicular tissue significantly improves sperm retrieval rates in non-obstructive azoospermia (NOA) patients. Enzymatic digestion and staining enhance sperm detection when initial methods fail, aiding fertility treatments.
Area of Science:
- Reproductive Medicine
- Andrology
- Laboratory Science
Background:
- Sperm extraction techniques like TESA and micro-TESE focus on surgical factors, often neglecting crucial laboratory processing of testicular tissue.
- Standardized laboratory procedures are lacking for non-obstructive azoospermia (NOA) patients, impacting the success of sperm retrieval.
- Optimizing laboratory protocols is essential for maximizing sperm yield in NOA cases.
Purpose of the Study:
- To develop and validate a standardized three-step laboratory processing method for testicular tissue.
- To assess the effectiveness of enzymatic digestion and staining in detecting spermatozoa in NOA patients.
- To identify predictors for successful sperm identification post-laboratory processing.
Main Methods:
- Enrolled 811 patients undergoing TESA or micro-TESE between July 2023 and December 2024.
- Analyzed 466 obstructive azoospermia and 345 non-obstructive azoospermia (NOA) cases.
- Evaluated sperm retrieval rate (SRR) and sperm detection rate (SDR) after mechanical mincing, enzymatic digestion, and staining.
Main Results:
- Initial mincing yielded spermatozoa in 79.9% of TESA and 32.5% of micro-TESE patients.
- Enzymatic digestion and staining identified spermatozoa in an additional 23.21% of cases where initial mincing failed.
- Seminiferous tubule diameter was the sole predictor for sperm identification in NOA patients after enzymatic digestion and staining.
Conclusions:
- A three-step laboratory processing method involving enzymatic digestion and staining significantly enhances sperm retrieval in NOA patients.
- Enzymatic digestion improves the sperm retrieval rate (SRR), while staining increases the sperm detection rate (SDR).
- Seminiferous tubule diameter serves as a valuable intraoperative predictor for sperm retrieval success.
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