title: Detrimental Effect of Phenol Red on the Vitrification of Cat (Felis catus) Ovarian Tissue.
abstract: The aim of this study was to investigate the effects of different media with or without phenol red or the antioxidant trolox on the successful vitrification of feline ovarian tissue. In a first experiment, ovarian cortical pieces from three cats were vitrified in solutions of Roswell Park Memorial Institute (RPMI)-1640 medium, Minimum Essential Medium, Dulbecco's modified Eagle's medium, or Tissue Culture Medium 199 as basic medium, supplemented or not with 50 μM of trolox, all containing 40% ethylene glycol (EG) and 1 M of sucrose. RPMI-1640 (phenol red-free) without trolox was the only medium that preserved the percentage of morphologically normal preantral follicles similar to control (80%). The main difference between RPMI-1640 and the other media was the absence of phenol red and CaCl2. In a second experiment, ovarian cortical pieces from three cats were vitrified in a solution containing RPMI-1640 as basic medium, 40% EG, 1 M of sucrose, supplemented or not with phenol red or CaCl2 alone, or in combination. It was observed that phenol red supplementation led to follicular degeneration. Finally, to evaluate the interaction between phenol red and the cryoprotectant agent (i.e., EG), ovarian tissue was exposed to RPMI-1640 supplemented with phenol red and EG at different concentrations (10%, 20%, or 40%). There was an inverse relationship between EG concentration and free phenol red in the medium after exposure. It is suggested that vitrification of feline ovarian tissue should be performed in a phenol red-free medium. Medium supplementation with 50 μM of trolox was deleterious for follicular morphology.
authors.affiliation: 1 Laboratory of Cytogenetics, Biological Sciences Institute, Federal University of Pará , Belem, Brazil .|2 Laboratory of Wild Animal Biology and Medicine, Federal University of Pará , Belem, Brazil .
authors.forename: Danielle C
authors.initials: DC
authors.lastname: Brito
authors.affiliation: 2 Laboratory of Wild Animal Biology and Medicine, Federal University of Pará , Belem, Brazil .
authors.forename: Sheyla F S
authors.initials: SF
authors.lastname: Domingues
authors.affiliation: 3 Laboratory Engineering of Natural Products, Federal University of Pará , Belem, Brazil .
authors.forename: Joyce K
authors.initials: JK
authors.lastname: Silva
authors.affiliation: 4 Faculty of Veterinary Medicine, Utrecht University , Utrecht, the Netherlands .
authors.forename: Xueqing
authors.initials: X
authors.lastname: Wu
authors.affiliation: 2 Laboratory of Wild Animal Biology and Medicine, Federal University of Pará , Belem, Brazil .|4 Faculty of Veterinary Medicine, Utrecht University , Utrecht, the Netherlands .
authors.forename: Regiane R
authors.initials: RR
authors.lastname: Santos
authors.affiliation: 1 Laboratory of Cytogenetics, Biological Sciences Institute, Federal University of Pará , Belem, Brazil .
authors.forename: Julio C
authors.initials: JC
authors.lastname: Pieczarka
mesh_terms: D000818:Animals; D002415:Cats; D003451:Cryoprotective Agents / Q000009:adverse effects*; D005260:Female; D010053:Ovary / Q000187:drug effects*; D010637:Phenolsulfonphthalein / Q000009:adverse effects*; D011309:Preservation, Biological / Q000379:methods*; D058989:Vitrification / Q000187:drug effects*
chemical_list: D003451:Cryoprotective Agents; D010637:Phenolsulfonphthalein