1. Cryopreservation of adult bovine testicular tissue for spermatogonia enrichment.
- Author
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Wu JJ, Hu TJ, Guo B, Yue ZP, Yang ZT, and Zhang XM
- Subjects
- Animals, Cattle, Cell Survival drug effects, Cryoprotective Agents pharmacology, Dimethyl Sulfoxide pharmacology, Epithelial Cells cytology, Epithelial Cells drug effects, Ethylene Glycol pharmacology, Freezing, Male, Propylene Glycol pharmacology, Seminiferous Epithelium cytology, Seminiferous Epithelium drug effects, Spermatogonia cytology, Spermatogonia drug effects, Testis cytology, Testis drug effects, Cryopreservation methods, Epithelial Cells physiology, Semen Preservation methods, Seminiferous Epithelium physiology, Spermatogonia physiology, Testis physiology
- Abstract
To develop a procedure for cryopreservation of adult bovine testis tissue, the effects of dimethyl sulphoxide (DMSO), propylene glycol (PG), ethylene glycol (EG), and their concentrations (v/v), as well as different thawing temperatures, on the cell viability of bovine testis tissue after freezing/thawing were examined. The highest testicular cell viabilities came from the media containing DMSO (85.3 ± 1.2 percent), PG (82 ± 1.0 percent) and EG (83.4 ± 1.0 percent) at 10 percent concentration respectively. Using 10 percent DMSO gave significantly higher spermatogonia percentage (61.1 ± 1.2 percent, P < 0.001) than processing with 10 percent PG (54.3 ± 0.6 percent) or 10 percent EG (55 ± 1.8 percent) after differential plating. Thawing in water bath of 37 or 97-100 degree C also provided significantly higher viabilities (85.1 ± 1.0, 85 ± 1.0 percent, P < 0.01, respectively) and spermatogonia percentages (56.6 ± 2.0, 56.6 ± 2.6 percent, P < 0.01, respectively) than that thawing at 4C (23.4 ± 0.8 percent for total viability, 8.97 ± 1.0 percent for spermatogonia percentage). Collectively, 10 percent DMSO and thawing in 37-100 degree C water baths were appropriate for the cryopreservation of bovine testicular tissue and subsequent spermatogonia enrichment.
- Published
- 2011