title: Freezing solution containing dimethylsulfoxide and fetal calf serum maintains survival and ultrastructure of goat preantral follicles after cryopreservation and in vitro culture of ovarian tissue.
abstract: Goat ovarian cortex fragments were subjected to slow freezing in the presence of various solutions containing intracellular cryoprotectants, including 1.0 M ethylene glycol (EG), propanediol (PROH), or dimethyl sulfoxide (DMSO), with or without sucrose and/or fetal calf serum (FCS). Histological examination revealed that only the DMSO-containing solutions were able to maintain a follicular ultrastructure similar to the morphology observed in the fresh control. Therefore, fragments previously cryopreserved in DMSO solutions (with and without sucrose and/or FCS) were cultured in vitro for 48 h and then subjected to viability, histological, and ultrastructural analysis. No significant differences were observed among the percentages of morphologically normal follicles in cryopreserved ovarian tissue before in vitro culture (DMSO: 62.5%; DMSO + sucrose: 68.3%; DMSO + FCS: 60.0%; DMSO + sucrose + FCS: 60.0%) and after culture (DMSO: 60.8%; DMSO + sucrose: 64.2%; DMSO + FCS: 70.8%; DMSO + sucrose + FCS: 55.0%). Following in vitro culture, the viability analysis showed that only the freezing solution containing DMSO and FCS (75.6%) maintained a percentage of viable follicles similar to that observed after culture without cryopreservation (89.3%). As determined by ultrastructural analysis, morphologically normal preantral follicles were detected in the fresh control and in fragments cultured before and after cryopreservation with DMSO and FCS. Thus, a freezing solution containing DMSO and FCS, under the experimental conditions tested here, guaranteed the maintenance of viability and follicular ultrastructure after short-term in vitro culture.
authors: Laboratory of Manipulation of Oocytes and Preantral Follicles, Faculty of Veterinary Medicine, State University of Ceará, Fortaleza, CE, Brazil. simone_medvet@hotmail.com
authors: Simone Vieira
authors: SV
authors: Castro
authors: Adeline Andrade
authors: AA
authors: de Carvalho
authors: Cleidson Manoel Gomes
authors: CM
authors: da Silva
authors: Luciana Rocha
authors: LR
authors: Faustino
authors: Cláudio Cabral
authors: CC
authors: Campello
authors: Carolina Madeira
authors: CM
authors: Lucci
authors: Sônia Nair
authors: SN
authors: Báo
authors: José Ricardo
authors: JR
authors: de Figueiredo
authors: Ana Paula Ribeiro
authors: AP
authors: Rodrigues
mesh_terms: D000818:Animals; D015925:Cryopreservation / Q000379:methods*; D003451:Cryoprotective Agents / Q000494:pharmacology*; D004121:Dimethyl Sulfoxide / Q000494:pharmacology*; D005260:Female; D005615:Freezing; D006041:Goats; D006080:Ovarian Follicle / Q000166:cytology / Q000187:drug effects / Q000648:ultrastructure*; D044967:Serum / Q000378:metabolism*; D012996:Solutions; D046509:Tissue Culture Techniques / Q000379:methods*; D014022:Tissue Survival / Q000187:drug effects*
chemical_list: D003451:Cryoprotective Agents; D012996:Solutions; D004121:Dimethyl Sulfoxide