Overnight Culturing of Testicular Sperm may Improve Pregnancy Outcomes

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Overnight culturing of testicular sperm may improve pregnancy outcomes

Effect of in vitro testicular spermatozoa culture on pregnancy outcomes: an experience at a single university hospital

A retrospective study at Ajou University Hospital was carried out to evaluate if in-vitro culturing of testicular spermatozoa significantly altered pregnancy outcomes in TESE-ICSI (testicular sperm extraction-intracytoplasmic sperm injection) cycles.

In total 46 couples, who male partners were diagnosed with nonobstructive or obstructive azoospermia, underwent 83 TESE-ICSI cycles between 2001 and 2015. Sperm and oocyte retrieval was performed on the same day for 65 of the cycles (Group A), while the remaining 18 cycles (Group B) had sperm retrieval performed a day earlier, followed by culturing of the biopsied samples for 24 hours, before extraction and fertilization via ICSI.

Sydney IVF fertilization medium (Cook Medical, Bloomington, IN, USA) with 10% human serum albumin was used for both group’s, with Group B tissue specifically cultured at 37°C, 6% CO2, 5% O2 and pH controlled for 24 hours. Ovarian stimulation protocols (Long agonist, short GnRH antagonist, pure FSH, FSH + hMG), were optimized according to individual patient characteristics (age, diagnosis and ovarian reserve). Patient characteristics showed no significant difference between Group A and Group B patients, with women of similar age, basal FSH and AMH levels, and also total number of stimulation days. In males however baseline LH levels was significantly different between Group A and B (3.67 ±1.07 IU/L vs 4.56 ±1.24 IU/L).

Statistical analysis of the results confirmed that the mean number of oocytes and good-quality oocytes retrieved was not significantly different between the 2 groups however both the fertilization (72.4% ±32.1% vs 59.2% ±21.7%) and implantation (35.0% ±34.1% vs 14.0% ±21.5%) rates was significantly higher in Group B vs Group A. This lead to a significantly higher clinical pregnancy rate per cycle (80% vs 37.5%) and pregnancy rate per cycle (80% vs 39%) for Group B vs Group A.

The authors acknowledged that the small and uneven sample size of the groups made it difficult to extrapolate further statistical significance from the data, stating larger sample studies are required to confirm these findings.


  1. Small study size
  2. Significantly uneven sample size between Group A and B
  3. No data on sperm motility
  4. Pooling of results (different ovarian stimulation protocols can influence pregnancy outcomes)

Similar studies

Hosseini A, et al. (2017). Improvement of motility after culture of testicular spermatozoa: the effects of incubation timing and temperature. https://doi.org/10.21037/tau.2017.03.43

Levran D, et al. (2001). Timing of testicular sperm retrieval procedures and in vitro fertilization-intracytoplasmic sperm injection outcome. https://doi.org/10.1016/S0015-0282(01)01908-2

Hu Y, et al. (1999). Clinical application of intracytoplasmic sperm injection using in vitro cultured testicular spermatozoa obtained the day before egg retrieval. https://doi.org/10.1016/S0015-0282(99)00308-8

Balaban B, et al. (1999). In-vitro culture of spermatozoa induces motility and increases implantation and pregnancy rates after testicular sperm extraction and intracytoplasmic sperm injection. https://doi.org/10.1093/humrep/14.11.2808


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