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Comparative efficacies of six different media for cryopreservation of immature buffalo (Bubalus bubalis) calf testis

Devi, Lalitha, Makala, Himesh, Pothana, Lavanya, Nirmalkar, Khemlal, Goel, Sandeep
Reproduction, fertility, and development 2016 v.28 no.7 pp. 872-885
DNA damage, Leydig cells, RNA-binding proteins, actin, apoptosis, buffaloes, calves, cell viability, cryopreservation, cytochrome P-450, dairies, dimethyl sulfoxide, fetal bovine serum, freezing, germplasm conservation, males, mortality, sexual maturity, signal transduction, testosterone, tissues, transcription factors, vimentin
Buffalo calves have a high mortality rate (~80%) in commercial dairies and testis cryopreservation can provide a feasible option for the preservation of germplasm from immature males that die before attaining sexual maturity. The aim of the present study was to evaluate combinations of 10 or 20% dimethylsulfoxide (DMSO) with 0, 20 or 80% fetal bovine serum (FBS) for cryopreservation of immature buffalo testicular tissues, subjected to uncontrolled slow freezing. Tissues cryopreserved in 20% DMSO with 20% FBS (D20S20) showed total, tubular and interstitial cell viability, number of early apoptotic and DNA-damaged cells, surviving germ and proliferating cells and expression of testicular cell-specific proteins (POU class 5 homeobox (POU5F1), vimentin (VIM) and actin α2 (ACTA2)) similar to that of fresh cultured control (FCC; P>0.05). Expression of cytochrome P450, family 11, subfamily A (CYP11A1) protein and testosterone assay showed that only tissues cryopreserved in D20S20 had Leydig cells and secretory functions identical to that of FCC (P>0.05). High expression of superoxide dismutase2 (SOD2), cold-inducible RNA-binding protein (CIRBP) and RNA-binding motif protein3 (RBM3) proteins in cryopreserved tissues indicated involvement of cell signalling pathways regulating cellular protective mechanisms. Similarity in expression of pro-apoptosis proteins transcription factor tumour protein P53 (TP53) and BCL2-associated X protein (BAX) in D20S20 cryopreserved tissues to that of FCC (P>0.05) suggested lower apoptosis and DNA damage as key reasons for superior cryopreservation.