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Merck

Testicular tissue cryopreservation is the preferred method to preserve spermatogonial stem cells prior to transplantation.

Reproductive biomedicine online (2020-02-01)
Jaime Onofre, Prashant Kadam, Yoni Baert, Ellen Goossens
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Which cryopreservation method better protects reproductive potential: the cryopreservation of a testicular cell suspension (TCS) or the cryopreservation of testicular tissue (TET)? Two cryopreservation strategies for spermatogonial stem cells (SSCs) were compared in a mouse model: cryopreservation as TET or as TCS. Evaluated outcomes were number of viable cells after thawing, number and length of donor-derived colonies after spermatogonial stem cell transplantation (SSCT), number of litters, litter size and number of donor-derived pups after mating. Compared with cryopreserving TCS, cryopreservation of TET resulted in significantly higher numbers of viable cells after thawing (TET: 13.4ย โ€ฏร—โ€ฏย 104ย ยฑย 7.2ย โ€ฏร—โ€ฏย 104 versus TCS: 8.2ย โ€ฏร—โ€ฏย 104ย ยฑย 2.7ย โ€ฏร—โ€ฏย 104; Pโ€ฏ=โ€ฏ0.0002), more (TET: 47.6ย ยฑย 19.2 versus TCS: 18.5ย ยฑย 13.0; Pย =ย 0.0039) and longer (TET: 5.2ย ยฑย 1.0ย mm versus TCS: 2.7ย ยฑย 1.5ย mm; Pย =ย 0.0016) donor-derived colonies, and more donor-derived pups per litter (TET: 2.2ย ยฑย 0.2 versus TCS: 0.5ย ยฑย 0.1; Pย =ย 0.0008). Cryopreservation of TET is the preferred method to cryopreserve SSCs prior to SSCT in a mouse model.

MATERIALS
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Sigma-Aldrich
Collagenase from Clostridium histolyticum, suitable for release of physiologically active rat hepatocytes, Type IV, 0.5-5.0 FALGPA units/mg solid, โ‰ฅ125 CDU/mg solid
Sigma-Aldrich
Hydrogen peroxide solution, contains inhibitor, 30 wt. % in H2O, meets USP testing specifications
Sigma-Aldrich
Ethylenediaminetetraacetic acid tetrasodium salt dihydrate, BioReagent, suitable for cell culture, 98.5-102.0%
Sigma-Aldrich
Formaldehyde solution, for molecular biology, BioReagent, โ‰ฅ36.0% in H2O (T)
Sigma-Aldrich
Deoxyribonuclease I from bovine pancreas, lyophilized powder, Protein ≥85 %, ≥400 Kunitz units/mg protein