Isolation, in vitro culture, and characterization of black crappie, Pomoxis nigromaculatus and white crappie, P. annularis ovarian tissue primary cells
Rattachement africain : us. Niveau de preuve : code pays fourni par la source.
Le résumé fourni par la source
Abstract Investigations were conducted to develop a general protocol for the isolation and in vitro culture of ovary‐derived cells from black crappie, Pomoxis nigromaculatus, and white crappie, P. annularis. Five digestive enzymes: 500 U/mL collagenase type I, 500 U/mL collagenase type IV, 0.05% trypsin‐EDTA, 0.25% trypsin‐EDTA, and 1X TrypLE™ Express were evaluated for live cell isolation. The isolated cells were cultured in 10 or 20% fetal bovine serum (FBS) in L‐15 growth media. In addition, four incubation temperatures (15°C, 20°C, 25°C, and 30°C) were also evaluated. The number of live cells obtained from the 0.25% trypsin and TrypLE™ Express treatments was significantly higher than other treatments. No difference in cell growth was observed between the two FBS treatments. Cells isolated using TrypLE™ Express and 0.25% trypsin reached 80% to 90% confluency in 12.5 cm2 cell culture flasks within 5 days of inoculation at 20 and 25°C. At 15°C, 10 days were required to reach 80%–90% confluency. Morphologically, cells incubated at 15°C, 20°C, and 25°C appeared healthier than cells incubated at 30°C, where irregular cell shape and substrate detachment were observed. We concluded TrypLE™ Express and 0.25% trypsin were optimal for cell dissociation and isolation of crappie ovarian cells. An incubation temperature of 20°C–25°C was favorable for cell culture in L‐15 media supplemented with either 10% or 20% FBS.
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Le contrôle bibliographique ouvert
DOI retrouvé dans Crossref DOI retrouvé, mais le titre doit être comparé manuellement.
- Titre Crossref
- Isolation, in vitro culture, and characterization of black crappie, <i>Pomoxis nigromaculatus</i> and white crappie, <i>P. annularis</i> ovarian tissue primary cells
- Date Crossref
- 10/05/2023
- Éditeur
- Wiley
- Type
- journal-article
Ce recoupement confirme des métadonnées liées au DOI. Il ne confirme ni la méthode ni les conclusions de l’étude, et il ne compte pas comme une seconde source scientifique indépendante.
Où se fait cette recherche
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University of Arkansas at Pine Bluff Department of Aquaculture and Fisheries pays non établi dans la noticeUniversité ou école supérieure
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Arkansas Arts Council pays non établi dans la noticeOrganisme public
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Arkansas Department of Agriculture pays non établi dans la noticeOrganisme public
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Auburn University pays non établi dans la noticeUniversité ou école supérieure
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Aquaculture Division pays non établi dans la noticeInstitution
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School of Fisheries pays non établi dans la noticeUniversité ou école supérieure
Department of Aquaculture and Fisheries — University of Arkansas at Pine Bluff, Arkansas Arts Council et Arkansas Department of Agriculture, avec 3 autres affiliations.
Une affiliation ne permet pas de déduire la nationalité d’un auteur.