Cryopreservation of adult bovine testicular tissue for spermatogonia enrichment.
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.
Department of Laboratory Medicine, Jilin University, Changchun, China.
J J
JJ
Wu
T J
TJ
Hu
B
B
Guo
Z P
ZP
Yue
Z T
ZT
Yang
X M
XM
Zhang
D000818:Animals; D002417:Cattle; D002470:Cell Survival / Q000187:drug effects; D015925:Cryopreservation / Q000379:methods*; D003451:Cryoprotective Agents / Q000494:pharmacology; D004121:Dimethyl Sulfoxide / Q000494:pharmacology; D004847:Epithelial Cells / Q000166:cytology / Q000187:drug effects / Q000502:physiology*; D019855:Ethylene Glycol / Q000494:pharmacology; D005615:Freezing; D008297:Male; D019946:Propylene Glycol / Q000494:pharmacology; D012662:Semen Preservation / Q000379:methods*; D012670:Seminiferous Epithelium / Q000166:cytology / Q000187:drug effects / Q000502:physiology*; D013093:Spermatogonia / Q000166:cytology / Q000187:drug effects / Q000502:physiology*; D013737:Testis / Q000166:cytology / Q000187:drug effects / Q000502:physiology*
D003451:Cryoprotective Agents; D019946:Propylene Glycol; D019855:Ethylene Glycol; D004121:Dimethyl Sulfoxide