What is the best protocol to cryopreserve immature mouse testicular cell suspensions?

Reproductive Biomedicine Online
Jaime OnofreEllen Goossens

Abstract

From a clinical perspective, which parameters grant optimal cryopreservation of mouse testicular cell suspensions? We studied the effect of different cryopreservation rates, the addition of sugars, different vessels and the addition of an apoptotic inhibitor on the efficiency of testicular cell suspension cryopreservation. After thawing and warming, testicular cell suspensions were transplanted to recipient mice for further functional assay. After selecting the optimal cryopreservation procedure, a second experiment compared the transplantation efficiency between the selected freezing protocol and fresh testicular cell suspensions. Multiple- and single-step freezing did not differ significantly in terms of recovered viable cells (RVC) (33 ± 28% and 38 ± 25%). The addition of sucrose did not result in a higher RVC (33 ± 20%). Cells frozen in vials recovered better than those frozen in straws (52 ± 20% versus 33 ± 20%; P = 0.0049). The inclusion of an apoptosis inhibitor (z-VAD[Oe]-FMK) significantly increased the RVC after thawing (61 ± 18% versus 50 ± 17%; P = 0.0480). When comparing the optimal cryopreservation procedure with fresh testicular cell suspensions, a lower RVC (63 ± 11% versus 92 ± 4%; P < 0.0001) and number of don...Continue Reading

Citations

Jan 10, 2021·In Vitro Cellular & Developmental Biology. Animal·J V MedranoE Novella-Maestre
Apr 2, 2021·Cell and Tissue Research·Tanushree PatraMukesh Kumar Gupta

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