title: Cryopreservation of ovine primordial follicles using dimethyl sulfoxide.
abstract: 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.
authors.affiliation: Laboratorio Renzo Giuliani, Dipartimento di Scienze Zootecniche, University of Florence, Florence, Italy. amorimc-kleinj@yahoo.com
authors.forename: Christiani Andrade
authors.initials: CA
authors.lastname: Amorim
authors.forename: Ana Paula Ribeiro
authors.initials: AP
authors.lastname: Rodrigues
authors.forename: Davide
authors.initials: D
authors.lastname: Rondina
authors.forename: Paulo Bayard Dias
authors.initials: PB
authors.lastname: Gonçalves
authors.forename: José Ricardo
authors.initials: JR
authors.lastname: de Figueiredo
authors.forename: Alessandro
authors.initials: A
authors.lastname: Giorgetti
mesh_terms: 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
chemical_list: D004396:Coloring Agents; D003451:Cryoprotective Agents; D014343:Trypan Blue; D004121:Dimethyl Sulfoxide