Validation-verification of a Highly Effective, Practical Human Testicular Tissue in Vitro Culture-cryopreservation Procedure Aimed to Optimize Pre-freeze and Post-thaw Motility

Overview

Journal J Assist Reprod Genet

Publisher Springer

Specialties Genetics
Reproductive Medicine

Date 2016 Feb 6

PMID 26847133

Citations 7

Authors

M C Schiewe

C Rothman

A Spitz

P E Werthman

S I Zeitlin

R E Anderson

Affiliations

Soon will be listed here.

Abstract

Purpose: 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.

Methods: 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.

Results: 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.

Conclusions: 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.

Citing Articles

Transcriptomic Differences by RNA Sequencing for Evaluation of New Method for Long-Time In Vitro Culture of Cryopreserved Testicular Tissue for Oncologic Patients.

Pei C, Todorov P, Kong Q, Cao M, Isachenko E, Rahimi G Cells. 2024; 13(18.

PMID: 39329723 PMC: 11430757. DOI: 10.3390/cells13181539.

Comparative Transcriptomic Analyses for the Optimization of Thawing Regimes during Conventional Cryopreservation of Mature and Immature Human Testicular Tissue.

Pei C, Todorov P, Cao M, Kong Q, Isachenko E, Rahimi G Int J Mol Sci. 2024; 25(1).

PMID: 38203385 PMC: 10778995. DOI: 10.3390/ijms25010214.

Sperm Selection Procedures for Optimizing the Outcome of ICSI in Patients with NOA.

Aydos K, Aydos O J Clin Med. 2021; 10(12).

PMID: 34207121 PMC: 8234729. DOI: 10.3390/jcm10122687.

Letter to Editor: We Need Standardized Guidelines for Laboratory Tissue Processing After Microsurgical Testicular Sperm Extraction.

Salama N Reprod Sci. 2021; 28(8):2071-2075.

PMID: 33871827 DOI: 10.1007/s43032-021-00585-4.

The evolution of testicular sperm extraction and preservation techniques.

Godart E, Turek P Fac Rev. 2021; 9:2.

PMID: 33659934 PMC: 7886077. DOI: 10.12703/b/9-2.

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