title: Ultrastructural features of agouti (Dasyprocta aguti) preantral follicles cryopreserved using dimethyl sulfoxide, ethylene glycol and propanediol.
abstract: The objective was to develop an efficient protocol for cryopreservation of agouti (Dasyprocta aguti) ovarian tissue. Agouti ovarian fragments were placed, for 10 min, in a solution containing MEM and fetal bovine serum plus 1.5 M dimethyl sulfoxide (DMSO), ethylene glycol (EG) or propanediol (PROH); some of those fragments were subsequently cryopreserved in a programmable freezer. After exposure and/or thawing, all samples were fixed in Carnoy prior to histological analysis. To evaluate ultrastructure, follicles from the control and all cryopreserved treatments were fixed in Karnovsky and processed for transmission electron microscopy. After exposure and freezing, there was a significant decrease in the percentage of morphologically normal preantral follicles in all treatments when compared to the control (92.67 ± 2.79, mean ± SD). However, there were no significant difference when the exposure and freezing procedures were compared using the same cryoprotectant. Moreover, there was no significant difference among cryoprotectants at the time of exposure (DMSO: 64.7 ± 3.8; EG: 70.7 ± 11.2, PROH: 63.3 ± 8.5) or after freezing (DMSO: 60.6 ± 3.6, EG: 64.0 ± 11.9; PROH: 62.0 ± 6.9). However, only follicles frozen with PROH had normal ultrastructure. In conclusion, preantral follicles enclosed in agouti ovarian tissue were successfully cryopreserved using 1.5 M PROH, with satisfactory maintenance of follicle morphology and ultrastructure.
authors.affiliation: Laboratory of Manipulation of Oocytes and Preantral Follicles (LAMOFOPA), Faculty of Veterinary, State University of Ceará, Fortaleza-CE, Brazil. livia_sch@yahoo.com.br
authors.forename: Livia Schell
authors.initials: LS
authors.lastname: Wanderley
authors.forename: Hiédely Kenia Machado
authors.initials: HK
authors.lastname: Luz
authors.forename: Luciana Rocha
authors.initials: LR
authors.lastname: Faustino
authors.forename: Isadora Machado Teixeira
authors.initials: IM
authors.lastname: Lima
authors.forename: Cláudio Afonso Pinho
authors.initials: CA
authors.lastname: Lopes
authors.forename: Alexandre Rodrigues
authors.initials: AR
authors.lastname: Silva
authors.forename: Sônia Nair
authors.initials: SN
authors.lastname: Báo
authors.forename: Claudio Cabral
authors.initials: CC
authors.lastname: Campello
authors.forename: Ana Paula Ribeiro
authors.initials: AP
authors.lastname: Rodrigues
authors.forename: José Ricardo
authors.initials: JR
authors.lastname: de Figueiredo
mesh_terms: D000818:Animals; D015925:Cryopreservation / Q000379:methods / Q000662:veterinary*; D004121:Dimethyl Sulfoxide; D056727:Endangered Species; D019855:Ethylene Glycol; D005260:Female; D006309:Hearing; D046529:Microscopy, Electron, Transmission / Q000662:veterinary; D023421:Models, Animal; D006080:Ovarian Follicle / Q000502:physiology / Q000648:ultrastructure*; D011409:Propylene Glycols; D012377:Rodentia / Q000033:anatomy & histology*; D014021:Tissue Preservation / Q000379:methods / Q000662:veterinary*
chemical_list: D011409:Propylene Glycols; D019855:Ethylene Glycol; D004121:Dimethyl Sulfoxide