BIOCHEMICAL AND ELECTROPHYSIOLOGICAL DIFFERENTIATION PROFILE OF A HUMAN NEUROBLASTOMA (IMR-32) CELL LINE

• Published in: In vitro cellular & developmental biology. Animal 2002-09, Vol.38 (8), p.450-456
• Main Authors: RAO, RAJ R, KISAALITA, WILLIAM S
• Format: Article
• Language: English
• Subjects: CELL AND TISSUE MODELS; Cell Differentiation; Cell lines; Cellular differentiation; cellular engineering; confocal microscopy; Cultured cells; Cytometry; Dyes; Flow Cytometry; Fluorescence; HEK293 cells; Humans; Membrane potential; Neuroblastoma; Neuroblastoma - enzymology; Neuroblastoma - metabolism; Neuroblastoma - pathology; Neuroblastoma - physiopathology; neuron-specific enolase; Neurons; Phosphopyruvate Hydratase - metabolism; resting membrane potential; Tumor Cells, Cultured
• ISSN: 1071-2690; 1543-706X; 1543-706X
• DOI: 10.1290/1071-2690(2002)038<0450:BAEDPO>2.0.CO;2

A human neuroblastoma cell line (IMR-32), when differentiated, mimics large projections of the human cerebral cortex and under certain tissue culture conditions, forms intracellular fibrillary material, commonly observed in brains of patients affected with Alzheimer's disease. Our purpose is to use differentiated IMR-32 cells as an in vitro system for magnetic field exposure studies. We have previously studied in vitro differentiation of murine neuroblastoma (N1E-115) cells with respect to resting membrane potential development. The purpose of this study was to extend our investigation to IMR-32 cells. Electrophysiological (resting membrane potential, Vm) and biochemical (neuron-specific enolase activity [NSE]) measurements were taken every 2 d for a period of 16 d. A voltage-sensitive oxonol dye together with flow cytometry was used to measure relative changes in Vm. To rule out any effect due to mechanical cell detachment, Vm was indirectly measured by using a slow potentiometric dye (tetramethylrhodamine methyl ester) together with confocal digital imaging microscopy. Neuron-specific enolase activity was measured by following the production of phosphoenolpyruvate from 2-phospho-d-glycerate at 240 nm. Our results indicate that in IMR-32, in vitro differentiation as characterized by an increase in NSE activity is not accompanied by resting membrane potential development. This finding suggests that pathways for morphological–biochemical and electrophysiological differentiations in IMR-32 cells are independent of one another.