Cryopreservation of ovine primordial follicles using dimethyl sulfoxide.
OBJECTIVE
To verify the viability of isolated primordial follicles after exposure to different concentrations of dimethyl sulfoxide (DMSO) and after cryopreservation.


METHODS
Randomized control trial.


METHODS
Laboratorio Renzo Giuliani, University of Florence, Italy.


METHODS
Thirty- to 40-day-old lambs.


METHODS
Isolated primordial follicles were stained with trypan blue to evaluate the effect of different DMSO concentrations before and after the cryopreservation. Histological structure and follicular mortality were evaluated.


RESULTS
After the isolation procedure (control), a mean (+/-SE) of 800 +/- 203.86 live primordial follicles/mL were obtained. The number of live follicles in the toxicity test using the DMSO at 0, 0.5, 1.0, 1.5, 2.0, and 2.5 M was 782 +/- 193.96, 754 +/- 172.03, 778 +/- 191.58, 736 +/- 191.92, 476 +/- 122.9, and 316 +/- 83.52, respectively. The number of live follicles at 2.5 M was lower than that in the control procedure. After cryopreservation, the numbers decreased to 0 +/- 0, 232 +/- 44.20, 636 +/- 161.82, 628 +/- 181.28, 208 +/- 11.57, and 184 +/- 47.07, respectively at 0, 0.5, 1.0, 1.5, 2.0, and 2.5 M. The number of live follicles at 0, 0.5, 2.0, and 2.5 M were lower than that in the control procedure.


CONCLUSIONS
After cryopreservation, only DMSO concentrations of 1.0 and 1.5 M showed a number of live follicles similar to that of the control procedure.
Amorim
Christiani Andrade
CA
Laboratorio Renzo Giuliani, Dipartimento di Scienze Zootecniche, University of Florence, Florence, Italy. amorimc-kleinj@yahoo.com
Rodrigues
Ana Paula Ribeiro
AP
Rondina
Davide
D
Gonçalves
Paulo Bayard Dias
PB
de Figueiredo
José Ricardo
JR
Giorgetti
Alessandro
A
D000818:Animals; D002470:Cell Survival / Q000502:physiology; D004396:Coloring Agents / Q000378:metabolism; D015925:Cryopreservation / Q000379:methods / Q000662:veterinary*; D003451:Cryoprotective Agents / Q000633:toxicity*; D004121:Dimethyl Sulfoxide / Q000633:toxicity*; D005260:Female; D006080:Ovarian Follicle / Q000187:drug effects*; D012756:Sheep / Q000502:physiology*; D014343:Trypan Blue / Q000378:metabolism
D004396:Coloring Agents; D003451:Cryoprotective Agents; D014343:Trypan Blue; D004121:Dimethyl Sulfoxide