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. 2004 Jan;44(1-2):15–25. doi: 10.1023/B:CYTO.0000043394.40309.48

Characterisation of Tetraploid and Diploid Clones of Spodoptera frugiperda Cell Line

RF Jarman-Smith, C Mannix, M Al-Rubeai
PMCID: PMC3449503  PMID: 19003226

Abstract

We have isolated and characterised diploid and tetraploid clones from the normally heterologous Spodoptera frugiperda (Sf-9)cell line by dilution cloning technique. Tetraploid clones were found to have cell sizes in excess of 35% larger than that of the diploid clones. In contrast, the maximum cell numbers achieved in batch cultures of diploid clones were on average 185% higher than the tetraploid cell numbers. Growth rates and metabolic quotients during the exponential phase were similar for both clones. Tetraploid cells infected with wild-type and recombinant green fluorescent protein (GFP) baculovirus, resulted in more polyhedra or GFP product per cell. Importantly, the difference between the clones either completely diminished or reduced to 50% when the yield was assessed in terms of the amount of polyhedra or GFP per mL of medium, respectively. These results indicate that the existing heterogeneity in insect cell populations with respect to ploidy level, are correlated to cell growth and product yield.

Keywords: AcNPV, Cell size, Flow cytometry, GFP, Insect cells, Intracellular pH, Spodoptera frugiperda

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