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Effects of various cryoprotectants on the quality of frozen–thawed immature bovine ( Qinchuan cattle ) calf testicular tissue
Author(s) -
Zhang X.G.,
Li H.,
Hu J.H.
Publication year - 2017
Publication title -
andrologia
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.633
H-Index - 59
eISSN - 1439-0272
pISSN - 0303-4569
DOI - 10.1111/and.12743
Subject(s) - cryoprotectant , cryopreservation , andrology , spermatogenesis , viability assay , biology , glycerol , trypan blue , gene expression , dimethyl sulfoxide , chemistry , cell , embryo , biochemistry , gene , medicine , microbiology and biotechnology , organic chemistry
Summary To investigate the effects of different concentrations of various cryoprotectants ( CP s) on the cell viability as well as expression of spermatogenesis‐related genes, such as CREM , Stra8 and HSP 70‐2 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 CP s: 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 HSP 70‐2 ) expression level came from the cryomedia containing glycerol, EG and DMSO at 10% concentration respectively ( p < .05). Meanwhile, compared with the other CP s, 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 HSP 70‐2 ).