title: Brilliant cresyl blue staining does not present cytotoxic effects on human luteinized follicular cells, according to gene/protein expression, as well as to cytotoxicity tests.
abstract: In vitro oocyte maturation presents many advantages and its success is related to the selection of fully grown oocytes. In animal models, staining of cumulus-oocyte complexes (COCs) with Brilliant Cresyl Blue (BCB) is widely used for this selection. However, a lack of information about the safety of BCB makes its applicability in humans questionable. Because granulosa and cumulus cells have a close relationship with the oocyte and taking into account that follicular cells are also exposed to BCB when this pre-selection method is applied, we aimed to evaluate the effects of BCB on human follicular cells exposed to BCB. Cytotoxicity tests (Sulforhodamine B and Neutral Red Uptake) and gene and protein expression of elements related to the cell cycle (BAX, BCL2, TP53 and CDKN1A), as well as to cell death and metabolism (GAPDH), glucose consumption, and estradiol and progesterone secretion, were examined in granulosa and cumulus cells with and without exposure to BCB. Regardless estradiol secretion and glucose consumption, all other evaluations presented similar results between control and treated (BCB) groups, independently of cell type. Therefore, our results demonstrate convincingly that BCB seems to be safe for use in humans and it should encourage future studies to evaluate the development of embryos derived from human oocytes selected by BCB staining.
authors.affiliation: Institute of Health Sciences/Department of Physiology, Universidade Federal do Rio Grande do Sul, 90050-170, Porto Alegre, Brazil. Electronic address: alcobadiego88@gmail.com.
authors.forename: Diego Duarte
authors.initials: DD
authors.lastname: Alcoba
authors.affiliation: Institute of Health Sciences/Department of Physiology, Universidade Federal do Rio Grande do Sul, 90050-170, Porto Alegre, Brazil.
authors.forename: Júlia
authors.initials: J
authors.lastname: Schneider
authors.affiliation: Institute of Health Sciences/Department of Physiology, Universidade Federal do Rio Grande do Sul, 90050-170, Porto Alegre, Brazil.
authors.forename: Letícia
authors.initials: L
authors.lastname: Arruda
authors.affiliation: Institute of Health Sciences/Department of Physiology, Universidade Federal do Rio Grande do Sul, 90050-170, Porto Alegre, Brazil.
authors.forename: Patrícia Borba
authors.initials: PB
authors.lastname: Martiny
authors.affiliation: Service of Gynecology and Obstetrics, Hospital de Clínicas de Porto Alegre, 90035-903, Porto Alegre, Brazil; Department of Gynecological Endocrinology and Reproductive Medicine, University Women's Hospital, 69120, Heidelberg, Germany.
authors.forename: Edison
authors.initials: E
authors.lastname: Capp
authors.affiliation: Institute of Health Sciences/Department of Physiology, Universidade Federal do Rio Grande do Sul, 90050-170, Porto Alegre, Brazil; Service of Gynecology and Obstetrics, Hospital de Clínicas de Porto Alegre, 90035-903, Porto Alegre, Brazil.
authors.forename: Helena
authors.initials: H
authors.lastname: von Eye Corleta
authors.affiliation: Institute of Health Sciences/Department of Physiology, Universidade Federal do Rio Grande do Sul, 90050-170, Porto Alegre, Brazil.
authors.forename: Ilma Simoni
authors.initials: IS
authors.lastname: Brum
mesh_terms: D000328:Adult; D018797:Cell Cycle Proteins / Q000235:genetics / Q000378:metabolism; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D004396:Coloring Agents / Q000633:toxicity*; D054885:Cumulus Cells / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D005191:Family Characteristics; D005260:Female; D015571:Follicular Fluid / Q000166:cytology; D018507:Gene Expression Regulation, Developmental / Q000187:drug effects*; D006107:Granulosa Cells / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D006801:Humans; D059471:In Vitro Oocyte Maturation Techniques; D007247:Infertility, Female / Q000378:metabolism / Q000473:pathology / Q000628:therapy; D007248:Infertility, Male; D034461:Luteinization; D008297:Male; D008875:Middle Aged; D054315:Oocyte Retrieval / Q000009:adverse effects; D006080:Ovarian Follicle / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D010078:Oxazines / Q000633:toxicity*
chemical_list: D018797:Cell Cycle Proteins; D004396:Coloring Agents; D010078:Oxazines; C006797:Brilliant Cresyl Blue
keywords: Brilliant cresyl blue (BCB); Cumulus cells; Cytotoxicity; Granulosa cells; In vitro maturation (IVM)