Graduate Thesis Or Dissertation
 

Partial characterization of six established salmonid cell lines

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  • Six salmonid cell lines previously established in this laboratory, CHSE 114 and CHSE 214 (chinook salmon), CSE 119 (coho salmon), SSE 5 (sockeye salmon), STE 137 (steelhead trout) and RTH 149 (rainbow trout), were partially characterized. Mycoplasmas were not detected in any of the cell lines. All cell lines exhibited heteroploidy. The CHSE 114, CHSE 214, and SSE 5 cell lines had hypodiploid modal chromosome numbers of 63, 67, and 64, respectively. The STE 137 and RTH 149 cell lines had hyperdiploid modal numbers of 66 and 64, respectively. The CSE 119 cell line had a hypotetraploid modal number of 110. The same modal number (67) was found at both transfer levels tested, 113 and 308, for CHSE 214 cell line. This indicated that the modal number for this cell line was no longer changing. Saturation density and optimum growth cell density were determined for all cell lines except CHSE 114. Saturation densities of cell lines ranged from 1.7 X 10⁶ (RTH 149) to 6.2 X 10⁶ (STE 137) cells per 35X10 mm dish. An optimum growth cell density of 2.3 X 10⁴ 5.0 X 10⁴, and 1.4 X 10⁴ cells per 35X10 mm dish was obtained for CHSE 214 (transfer level 100's), CSE 119, and STE 137 cell lines, respectively (t test, α .05). No differences in cell growth rates were found for other cell lines at densities tested (t test, α .05). Optimum growth temperature was determined for CHSE 214 and STE 137 cell lines grown at 8, 12, 16, 23, 27, and 34°C (t test, α .05). The CHSE 214 cell line exhibited exponential growth at 12, 16, and 23°C. The STE 137 cell line exhibited exponential growth at 12 and 16°C. Optimum growth temperatures for CHSE 214 cell line occurred at 16 and 23°C. No difference (α .05) between growth rates at 12 and 16°C was found for STE 137 cell line. Maintenance growth occurred at 8 and 27°C for CHSE 214 cell line, and at 8, 23, and 27°C for STE 137 cell line. Neither cell line grew at 34°C. No cells adhered to dishes after 72 h incubation at this temperature. Using a plaque assay technique, CHSE 114, CHSE 214, SSE 5, STE 137, and RTH 149 cell lines were susceptible to IHNV and IPNV. The CSE 119 cell line was found to be unsuitable for use in plaque assays. Both control and virus-infected CSE 119 cell monolayers developed plaques similar to those produced by viruses. Attempts at cloning CHSE 214, CSE 119, STE 137, and RTH 149 cell lines were futile. Cells would not grow at low densities required for cloning.
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file details PhilipponFriedMarie1980.CV01.pdf 2017-08-10 Pubblico Scaricare