title: Effects of various cryoprotectants on the quality of frozen-thawed immature bovine (Qinchuan cattle) calf testicular tissue.
abstract: To investigate the effects of different concentrations of various cryoprotectants (CPs) on the cell viability as well as expression of spermatogenesis-related genes, such as CREM, Stra8 and HSP70-2 in frozen-thawed bovine calf testicular tissue, immature bovine (Qinchuan cattle) calf testicular tissue was collected and cryopreserved in the cryomedia containing different concentrations (5%, 10%, 15% and 20%) of the following three CPs: glycerol, ethylene glycol (EG) and dimethyl sulphoxide (DMSO) respectively. After 1 month cryopreservation in liquid nitrogen, cell viability was evaluated using Trypan blue exclusion under a bright-field microscope. The mRNA expression of the three genes was also evaluated using qRT-PCR. The results indicated that different concentrations of glycerol, EG and DMSO in cryomedia during cryopreservation could protect bovine calf testicular tissue in various ways to avoid freezing or cryopreservation-induced expression changes in spermatogenesis-related genes. The highest cell viability and the three spermatogenesis-related genes (CREM, Stra8 and HSP70-2) expression level came from the cryomedia containing glycerol, EG and DMSO at 10% concentration respectively (p < .05). Meanwhile, compared with the other CPs, the frozen-thawed bovine calf testicular tissue treated with 10% DMSO exhibited the highest cell viability and mRNA expression level of the spermatogenesis-related genes (CREM, Stra8 and HSP70-2).
authors.affiliation: College of Animal Science and Technology, Northwest A&F University, Yangling, Shaanxi, China.
authors.forename: X-G
authors.initials: XG
authors.lastname: Zhang
authors.affiliation: College of Animal Science and Technology, Northwest A&F University, Yangling, Shaanxi, China.
authors.forename: H
authors.initials: H
authors.lastname: Li
authors.affiliation: College of Animal Science and Technology, Northwest A&F University, Yangling, Shaanxi, China.
authors.forename: J-H
authors.initials: JH
authors.lastname: Hu
mesh_terms: D000818:Animals; D002417:Cattle; D002470:Cell Survival; D015925:Cryopreservation; D003451:Cryoprotective Agents / Q000008:administration & dosage*; D051786:Cyclic AMP Response Element Modulator / Q000378:metabolism; D018840:HSP70 Heat-Shock Proteins / Q000378:metabolism; D008297:Male; D013737:Testis
chemical_list: D003451:Cryoprotective Agents; D018840:HSP70 Heat-Shock Proteins; D051786:Cyclic AMP Response Element Modulator
keywords: bovine calf testicular tissue; cell viability; cryopreservation; gene expression