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  • 1
    ISSN: 1573-0603
    Keywords: CHSE-214 ; H33258 ; Cell proliferation ; Calcium requirement ; CytoFluor 2350
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The use of a DNA-binding dye, bisbenzimidazole (H33258), and a microplate fluorometer, CytoFluor 2350, was optimized to measure cell number in Chinook salmon embryo cell cultures (CHSE-214). The uniformity of cell homogenates, which were prepared prior to staining, was evaluated by area scan, which consists of multiple measurements over the total well area. Disruption in 0.01% SDS produced a relatively uniform homogenate and a low background. Homogenates were stained with H33258 at 1 to 10 µg/ml. Linear relationships between cell numbers and fluorescence units were established in 96 well plates, which were read only by standard scan, and in 6, 12, 24 and 48 well plates, which were read by area and standard scan. Area scan provided better relationships. These methods were used to show that in L-15 medium CHSE-214 were able to attach, retain viability, and proliferate with very little exogenous calcium.
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    Springer
    Cytotechnology 16 (1994), S. 159-166 
    ISSN: 1573-0778
    Keywords: Fish cells ; glutamine-free growth ; nutrition
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract The glutamine requirement for thein vitro proliferation of fish cells was investigated with cell lines from four different species and three tissues: goldfish skin (GFSk-S1), Chinook salmon embryo (CHSE-214), and raibow trout liver (RTL-W1) and spleen (RTSp-W1). With a supplement of fetal bovine serum, the basal medium, Leibovitz's L-15, without glutamine supported the proliferation of all four cell lines as well, or nearly as well, as L-15 with 2 mM glutamine. This was true over short term assays of two to four weeks and for continuous propagation. CHSE-214 also grew as well with or without 2 mM glutamine in Minimum Essential Medium with fetal bovine serum. However, when the supplement was dialyzed fetal bovine serum, CHSE-214 grew much better in L-15 without glutamine. Therefore, glutamine was not required for growth in L-15, and in fact, was inhibitory in the absence of the dialyzable fraction of serum. By contrast, glutamine appeared to be important for growth in Minimum Essential Medium. When the supplement was dialyzed fetal bovine serum, CHSE-214 grew much better in Minimum Essential Medium with 2 mM glutamine. These results suggest that the glutamine requirement for thein vitro proliferation of fish cells is conditional and depends on the basal medium and serum supplement.
    Type of Medium: Electronic Resource
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