Cell cycle, cell size and mitochondrial activity of hybridoma cells during batch cultivation

Abstract

Cell cycle, cell size and rhodamine 123 fluorescence in cell populations of two batch cultures were analysed and quantified with a fluorescence-activated cell sorter (FACS). Two cultures derived from either exponential or stationary phase innocula were investigated in order to demonstrate the dependency of the subsequent cell growth on innoculum condition. The results demonstrated that the level of activity of cells in the innoculum culture could have a significant effect on cellular activity during the initial phase of the inoculated culture, as it advances through its growth cycle. Positive correlation was found between the cell size and mitochondrial activity (as measured by rhodamine 123 uptake) with S and G2 fractions as the cell progressed through the cell cycle. The enumeration of the fractions of cell cycle phases has helped in prediction of the changes in cell numbers following perturbation of the culture condition.

Uncoupling of cell growth and proliferation results in enhancement of productivity in p21CIP1‐arrested CHO cells

“…This therefore forms the physiological basis for enhanced productivity in arrested cells. G1-phase of the cell cycle of some cell lines may be a privileged position for increased production of heterologous proteins (Al-Rubeai and Emory, 1990;Al-Rubeai et al, 1991Kaufman et al, 2001;Lloyd et al, 2000;Suzuki and Ollis, 1990). At the start of the cell cycle, when cells are preparing for DNA replication and cell division, there is a requirement to replenish energy pools and metabolic precursors as well as increase cell mass (Ko and Prives, 1996;Linke et al, 1996).…”

Effect of feed and bleed rate on hybridoma cells in an acoustic perfusion bioreactor: Part I. Cell density, viability, and cell‐cycle distribution

“…The terms on the right side of the equal sign represent loss of cells due to the bleed and cell death, which is assumed to be independent of cell age. It is assumed for the moment that hybridoma cells do not go into quiescence (Al-Rubeai et al, 1991) and either grow or die. Under steady-state conditions the cell-age distribution does not change with time, and integration of Eq.…”

Online flow cytometry for monitoring apoptosis in mammalian cell cultures as an application for process analytical technology

“…These parameters have traditionally been restricted, due to technical challenges, to only a few, such as pH, dissolved oxygen, optical density, capacitance, and temperature (Carvell and Dowd 2006;Hanson et al 2007;Konstantinov et al 1992;Naciri et al 2008;Zeiser et al 1999). The limitation of these common measured parameters is that no direct information is given on the state of the cell such as viability, cell cycle, cell size, or apoptotic state, all of which are metrics that can help improve recombinant protein production and reproducibility of a mammalian culture bioprocess (Al-Rubeai et al 1991a;Ibarra et al 2003;Plasier et al 1999). Parameters that directly give cell and product quality information can improve upon current cell culture processes together with traditional parameters to provide dynamic modelling and prediction on product quantity and quality in real time and validate the bioprocess to the best extent (Streefland et al 2013).…”