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Titolo:
Updating quality control assays in the assisted reproductive technologies laboratory with a cryopreserved hamster oocyte DNA cytogenotoxic assay
Autore:
Chan, PJ; Calinisan, JH; Corselli, JU; Patton, WC; King, A;
Indirizzi:
Loma Linda Univ, Sch Med, Dept Gynecol & Obstet, Loma Linda, CA 92350 USA Loma Linda Univ Loma Linda CA USA 92350 Obstet, Loma Linda, CA 92350 USA
Titolo Testata:
JOURNAL OF ASSISTED REPRODUCTION AND GENETICS
fascicolo: 3, volume: 18, anno: 2001,
pagine: 129 - 134
SICI:
1058-0468(200103)18:3<129:UQCAIT>2.0.ZU;2-2
Fonte:
ISI
Lingua:
ENG
Soggetto:
MOUSE PREIMPLANTATION EMBRYOS; HUMAN INVITRO FERTILIZATION; COMET ASSAY; WATER-QUALITY; CULTURE; SPERM; CELL; SYSTEM; DAMAGE;
Keywords:
single-cell gel electrophoresis; comet assay; quality control; hamster ova; in vitro fertilization;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Clinical Medicine
Citazioni:
24
Recensione:
Indirizzi per estratti:
Indirizzo: Chan, PJ Loma Linda Univ, Sch Med, Dept Gynecol & Obstet, 11370 Anderson St,Suite 3950, Loma Linda, CA 92354 USA Loma Linda Univ 11370 Anderson St,Suite 3950 Loma Linda CA USA 92354
Citazione:
P.J. Chan et al., "Updating quality control assays in the assisted reproductive technologies laboratory with a cryopreserved hamster oocyte DNA cytogenotoxic assay", J AS REPROD, 18(3), 2001, pp. 129-134

Abstract

Purpose: Despite advances in assisted reproduction, there is no progress in (quality control bioassays. The objectives were to develop a comet assay to measure DNA fragmentation in thawed cryopreserved oocytes and compare this assay with one-cell mouse embryo bioassay. Methods: Thawed hamster oocytes from a commercial source were incubated inculture media with either 0-, 50-, or 100-muM hydrogen peroxide, or, in media exposed to different contact materials and unknown proficiency analytes. Incubation time was 1.5 h at 37 degreesC. The oocytes were dried filed, stained with acridine orange, embedded in a mini-agarose layer and electrophoresis was carried out. Fluorescent images were analyzed The results were compared with standard one-cell mouse assay data. Results: The 100-muM hydrogen peroxide treatment caused greatest DNA fragmentation in the hamster oocytes at Hours 1 and 2. A dose response was observed. Intrassay coefficient of variation was 5.7%. Only one of the five materials tested passed both assays The data for the unknown proficiency analytes were similar for both assaysConclusions: The oocyte comet assay demonstrated DNA fragmentation in the presence of toxic substances. The detection of toxicity in two materials that passed the mouse bioassay suggested increased sensitivity in the new assay The oocyte comet assay and the mouse bioassay results marched in the proficiency test. However, more studies are still needed to determine optimal sensitivity.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 05/07/20 alle ore 13:18:46