title: Comparative efficacies of six different media for cryopreservation of immature buffalo (Bubalus bubalis) calf testis.
abstract: 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.
authors.affiliation: Laboratory for the Conservation of Endangered Species, Centre for Cellular and Molecular Biology, Council for Scientific and Industrial Research, Uppal Road, Hyderabad, 500 007, India.
authors.forename: Lalitha
authors.initials: L
authors.lastname: Devi
authors.affiliation: Laboratory for the Conservation of Endangered Species, Centre for Cellular and Molecular Biology, Council for Scientific and Industrial Research, Uppal Road, Hyderabad, 500 007, India.
authors.forename: Himesh
authors.initials: H
authors.lastname: Makala
authors.affiliation: Laboratory for the Conservation of Endangered Species, Centre for Cellular and Molecular Biology, Council for Scientific and Industrial Research, Uppal Road, Hyderabad, 500 007, India.
authors.forename: Lavanya
authors.initials: L
authors.lastname: Pothana
authors.affiliation: Laboratory for the Conservation of Endangered Species, Centre for Cellular and Molecular Biology, Council for Scientific and Industrial Research, Uppal Road, Hyderabad, 500 007, India.
authors.forename: Khemlal
authors.initials: K
authors.lastname: Nirmalkar
authors.affiliation: Laboratory for the Conservation of Endangered Species, Centre for Cellular and Molecular Biology, Council for Scientific and Industrial Research, Uppal Road, Hyderabad, 500 007, India.
authors.forename: Sandeep
authors.initials: S
authors.lastname: Goel
mesh_terms: D000818:Animals; D002020:Buffaloes; D015925:Cryopreservation / Q000662:veterinary*; D003451:Cryoprotective Agents / Q000737:chemistry*; D004121:Dimethyl Sulfoxide / Q000737:chemistry; D005615:Freezing; D008297:Male; D013737:Testis / Q000502:physiology*
chemical_list: D003451:Cryoprotective Agents; D004121:Dimethyl Sulfoxide