title: Adding ascorbic acid to vitrification and IVC medium influences preantral follicle morphology, but not viability.
abstract: In this study, we analysed the effect on morphology and viability of ovine primordial follicles, when ascorbic acid (AA) was added to vitrification and in vitro culture (IVC) media. For morphological analysis, ovarian tissue was vitrified using DMSO or ethylene glycol (EG), to which AA was added or omitted. After warming, the tissue was fixed for histology or 1-day cultured in the presence or absence of AA. Isolated primordial follicles from ovine ovarian tissue vitrified with DMSO or EG, both supplemented with AA were stained with trypan blue for viability analysis, or 5-day cultured with or without AA followed by a viability analysis. In this study, we report on the successful vitrification protocol developed for ovine ovarian tissue using EG. Vitrification using DMSO reduced the percentage of morphological normal primordial follicles, whereas addition of AA to the vitrification and culture media did enhance these results (p < 0.05). However, vitrification in a DMSO + AA medium followed by 5-day IVC resulted in a significant decrease in the follicular viability, independently of the presence of AA in the IVC medium.
authors: Laboratory of Manipulation of Oocytes and Ovarian Preantral Follicles, Faculty of Veterinary, State of Ceará University, Fortaleza, CE, Brazil.
authors: M A P
authors: MA
authors: Melo
authors: I C
authors: IC
authors: Oskam
authors: J J H
authors: JJ
authors: Celestino
authors: A A
authors: AA
authors: Carvalho
authors: S V
authors: SV
authors: Castro
authors: J R
authors: JR
authors: Figueiredo
authors: A P R
authors: AP
authors: Rodrigues
authors: R R
authors: RR
authors: Santos
mesh_terms: D000818:Animals; D001205:Ascorbic Acid / Q000737:chemistry / Q000494:pharmacology*; D003451:Cryoprotective Agents / Q000737:chemistry / Q000494:pharmacology; D003470:Culture Media / Q000737:chemistry*; D005260:Female; D006080:Ovarian Follicle / Q000187:drug effects*; D012756:Sheep; D046509:Tissue Culture Techniques; D058989:Vitrification / Q000187:drug effects*
chemical_list: D003451:Cryoprotective Agents; D003470:Culture Media; D001205:Ascorbic Acid