Validation-verification of a highly effective, practical human testicular tissue in vitro culture-cryopreservation procedure aimed to optimize pre-freeze and post-thaw motility.
Schiewe. M C MC; Rothman. C C; Spitz. A A; Werthman. P E PE; Zeitlin. S I SI; Anderson. R E RE
Key Findings
- Incubating whole testicular biopsy pieces at 30 °C for 24‑96 hours before freezing preserves the highest percentage of motile sperm compared with 21 °C or 37 °C.
- Post‑thaw motility can reach up to 85 % of the pre‑freeze level, allowing easy selection of sperm for ICSI.
- Adding a sucrose dilution step after thawing restores sperm longevity to levels seen in fresh tissue and leads to successful embryo transfers.
Practical Outcomes
- For biohackers or DIY fertility enthusiasts, the protocol suggests that a short pre‑freeze incubation at 30 °C and freezing the tissue whole (without extensive processing) can dramatically improve sperm quality after thawing. However, because the research focuses on clinical testicular tissue and does not involve the peptide ovagen, it offers no direct actionable insight for peptide‑based self‑experiments.
Summary
The study describes a new way to handle and freeze whole pieces of testicular tissue so that sperm motility is kept high before freezing and after thawing. By incubating the tissue at 30 °C for 24‑96 hours before freezing, they kept up to 85 % of the sperm’s movement ability and got good results when the sperm were later used for ICSI. The method is simpler and requires less lab work than traditional approaches.
Abstract
The aim of our paper was to validate a testicular biopsy procedure that simplifies handling, processing, and cryopreservation, while at the same time optimizes sperm motility before freezing and after thawing. Two prospective studies were conducted to verify, optimize, and understand the virtues of pre-freeze testicular tissue IVC at different temperatures (21, 30, or 37 °C). Testicular tissue was obtained from clinical specimens designated for whole tissue cryopreservation (i.e., intact mass of tubules) and/or for fresh use in IVF-ICSI cycles. Whole testicular biopsy pieces (1-3 mm(3)) were diluted in glycerol containing freeze solutions, slow cooled to 4 °C and then rapidly frozen in LN2 vapor. Fresh and post-thaw testicular biopsy tissue were evaluated for changes in the quantity (%) and pattern of motility (I-IV: twitching to rapid progression, respectively) over a 1 week duration. The clinical effectiveness of IVC-cryopreserved whole testicular biopsy tissue was also validated analyzing fresh embryo transfers. More reliable recovery of motile testicular sperm was achieved using whole tissue freeze preservation combined with IVC (24-96 h) post-acquisition at an incubation temperature of 30 °C compared to ambient temperature (21 °C) or 37 °C. Up to 85 % of the pre-freeze motility was conserved post-thaw (+3 h) for easy ICSI selection. Sperm longevity was optimized to fresh tissue levels by implementing testicular biopsy sucrose dilution post-thaw. Favorable clinical outcomes were proven using frozen-thawed testicular biopsy sperm for ICSI. By employing minimal tissue manipulation, integrating pre-freeze IVC processing at 30 °C and the freezing of whole testicular biopsy tissue, we have reduced the labor and improved the efficacy of processing testicular tissue for freeze-preservation and subsequent ICSI use.
Study Information
pubmed
2016
2016-02-04T00:00:00.000Z
10.1007/s10815-016-0659-7
18
50