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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.