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- W4382489306 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." @default.
- W4382489306 created "2023-06-29" @default.
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- W4382489306 date "2023-05-10" @default.
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- W4382489306 title "Isolation, in vitro culture, and characterization of black crappie, <i>Pomoxis nigromaculatus</i> and white crappie, <i>P. annularis</i> ovarian tissue primary cells" @default.
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- W4382489306 doi "https://doi.org/10.1111/jwas.12981" @default.
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