Characterisation of Tetraploid and Diploid Clones of Spodoptera frugiperda Cell Line

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.

Genome Assembly and Annotation of the Trichoplusia ni Tni-FNL Insect Cell Line Enabled by Long-Read Technologies

“…In the context of the putative tetraploid state of Tni-FNL, these data are consistent with the previous identification of 28–30 chromosomes for most lepidopteran organisms. The chromosome number appears to be lower than previous studies on Sf9 cells, which suggests much higher levels of chromosome fragmentation in those cells [ 35 ].…”

“…Previous analysis of the chromosome content of lepidopteran cell lines showed a lack of consistency in the ploidy of these cells. Sf9 cell lines were shown to have a mixed population of diploid and tetraploid cells that varied in their ratios even when cells were cloned out [ 35 ]. Bombyx mori cell lines were mostly diploid but by examining a number of lepidopteran lines, ploidy was found to be highly variable [ 36 ].…”

Metabolic profiling of insect cell lines: Unveiling cell line determinants behind system's productivity

“…This work focused on the metabolic characterization of two commercial insect cell lines, Sf9 and High Five, which have achieved great popularity in the biotechnological field. Clonal selection and lab‐to‐lab variability of insect cell lines may influence several parameters, such as productivity, fluxes, and metabolite pools (Jarman‐Smith et al, ). Moreover, differences in cell size can hinder the productivity phenotype, since it is a direct function of the cell productive volume (Bédard et al, ; Taticek et al, ).…”