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Magnesium Content and Distribution During Differentiation of Hl60 Cells

Published online by Cambridge University Press:  02 July 2020

A. Di Francesco
Affiliation:
Department of Molecular Cardiology, Research Institute, The Cleveland Clinic Foundation, Cleveland, OH. Institute of General Pathology, Catholic University School of Medicine, Rome Italy
R.W. Desnoyer
Affiliation:
Department of Molecular Cardiology, Research Institute, The Cleveland Clinic Foundation, Cleveland, OH.
F.I. Wolf
Affiliation:
Institute of General Pathology, Catholic University School of Medicine, Rome Italy
V. Covacci
Affiliation:
Institute of General Pathology, Catholic University School of Medicine, Rome Italy
A. Cittadini
Affiliation:
Institute of General Pathology, Catholic University School of Medicine, Rome Italy
M. Bondt
Affiliation:
Department of Molecular Cardiology, Research Institute, The Cleveland Clinic Foundation, Cleveland, OH.
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Extract

Mg is present in all living cells as free and bound Mg. Previous observations have shown a positive correlation between total Mg content and proliferative rate in several different cell types (1). To investigate the association between Mg and cell proliferation, we utilized a human promyelocytic leukemia cell line, HL60, that can be induced to differentiate in vitro to neutrophil-like cells, by retinoic acid (RA) and other compounds.

To further investigate this question, we analyzed elemental compartmentalization in HL60 cells before and after differentiation using electron probe microanalysis (EPMA). Differentiation to neutrophil-like cells was induced by culturing the HL60 cells for 96 hours in the presence of 1 μM RA. Cells were washed, pelleted, then rapidly frozen in liquid ethane, cooled to -185°C with liquid N2. Ultrathin cryosections were cut from the surface of the frozen cells, freeze-dried overnight and carbon coated. X ray spectra were collected at -100°C and at 80 keV accelerating voltage in a Philips CM 12 STEM equipped with LaB6 gun and an EDAX 30-mm2 Si(Li) energy dispersive x-ray detector and multichannel analyzer.

Type
Quantitative Biological and Materials Microanalysis by Electrons and X-Rays
Copyright
Copyright © Microscopy Society of America 1997

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References

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