Starvation‐induced programmed death of hybridoma cells: Prevention by amino acid mixtures

Abstract

Association of the availability of nutrients with the phenomenon of programmed cell death—apoptosis—was investigated using hybridoma cells cultured in protein‐free medium under conditions of starvation, i.e., in RPMl‐1640 medium diluted to 50% with saline. Amino acid mixtures, such as MEM essential amino acids or MEM nonessential amino acids were found to prevent starvation death significantly when added to the diluted medium in 1 to 2 mM concentrations, the MEM vitamin mixture was ineffective, and glutamine displayed a moderate growth‐supporting effect. The specific monoclonal antibody production rate in cultures supplemented with amino acid mixtures was strikingly low, whereas supplementation with glutamine alone or simultaneously with other amino acids resulted in a specific antibody production rate comparable with the rate observed in undiluted medium. © 1995 John Wiley & Sons, Inc.

Reversible developmental stasis in response to nutrient availability in theXenopus laevisCNS

“…An increase in cell proliferation is often coupled with an increase in cell death (Herrup and Busser, 1995;Ross, 1996), yet lack of nutrients can also induce cell death (Bursch et al, 2008;Franek, 1995). To test whether NR or DF induced cell death, we counted the number of dying cells in the tectum using SytoxO labeling (Faulkner et al, 2015) in the three nutrition treatment groups.…”

Dose-dependent reduction of apoptosis in nutrient-limited cultures of NS/0 myeloma cells transfected with the E1B-19K adenoviral gene

“…Also, hybridomas and NS/0 myelomas cultivated in medium devoid of glutamine, glucose, cystine, or oxygen will rapidly die by apoptosis (Mercille and Massie, 1994a,b;Singh et al, 1994), whereas incubation in the presence of elevated levels of metabolic by-products such as ammonia and lactate induces necrotic cell death in these cells. The use of feeding strategies aimed at constantly supplying the cells with sufficient nutrients may considerably delay the induction of apoptotic cell death (Franek and Dolnikova, 1991;Duval et al, 1991;Robinson et al, 1994;Franek, 1995;Franek and ChladkovaSramkova, 1995;DiStephano et al, 1996). While such strategies may indeed improve productivity, they also involve retention of the protein of interest within the culture environment for extended periods of time.…”

Modulation of cell cycle progression and of antibody production in mouse hybridomas by a nucleotide analogue

“…We explain this fact by exhaustion of critical substrates, likely to occur in cultures run in the trivial batch mode. Therefore, in similar experiments carried out with the hybridoma PVA-187 the spinner cultures were fed with glutamine to ensure maximum energy supply for MAb synthesis (Franȇk, 1995;Franȇk andSrámková, 1998). The data of a representative experiment of this series are illustrated in Figure 3.…”

“…AI 50 -PMEG concentration at which the apoptotic index is equal to 50%. The experimental error of the data was ±10 and ±15% for cell number and for apoptotic index, respectively (Franȇk, 1995). protein production by mammalian cells (Takahashi et al, 1994).…”

“…Reduction of hybridoma growth by controlled feeding had, in our experience, a negative impact on the specific MAb production rate (Franȇk, 1995;Franȇk andSrámková, 1997, 1998. Therefore, we decided to pursue an alternative approach based on the introduction of an antiproliferative molecular signal in a culture system saturated with substrates.…”