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. 2005 Nov 30;46(2-3):97–107. doi: 10.1007/s10616-005-0301-5

Toxic Concentrations of Exogenously Supplied Methylglyoxal in Hybridoma Cell Culture

Benjamin M Roy 1,2, Tiffany D Rau 1, R Robert Balcarcel 1,✉
PMCID: PMC3449709  PMID: 19003265

Abstract

Concentrations at which methylglyoxal, a by-product of cellular metabolism, can be toxic to hybridoma cell cultures were determined using exogenously supplied doses. Trypan blue cell counts of 6-well cultures incubated for 24 h with various methylglyoxal concentrations revealed inhibition of cell growth at 300 μM and higher, with a median inhibitory concentration of 490±20 μM. The primary mode of death was apoptosis, as assessed by chromatin condensation, and the effects of methylglyoxal were observed to be complete by approximately eight hours. Yet, the impact of methylglyoxal was a function of the rate of dosing; stepwise addition of MG during the first 6 h of incubation inhibited growth but caused much less cell death than a comparable bolus dose. Inhibition of cellular metabolism by MG was found to coincide with inhibition of cell growth, with a comparable median inhibitory concentration of 360±20 μM. The effects on viable cell density and metabolism were both linear at doses approaching zero, with lowest observable effect levels of 54 and 77 μM, respectively. These results provide quantitative estimates for concentrations of methylglyoxal that may be inhibitory to biopharmaceutical-producing cell lines.

Keywords: Apoptosis, Hybridoma, Lactate, Methylglyoxal, Toxicity

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Glossary

AOEB

acridine orange/ethidium bromide

CHO

Chinese Hamster ovary

LOEL

lowest observable effect level

HSD

honest significant difference

IC50

median inhibitory concentration

IMDM

Iscove’s modified Dulbecco’s Medium

MG

methylglyoxal

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