A non-radioactive method for measuring Cu uptake in HepG2 cells

Fosset, C., McGaw, B. A., Reid, M. D. and McArdle, H. J. 2005, A non-radioactive method for measuring Cu uptake in HepG2 cells, Journal of inorganic biochemistry, vol. 99, no. 5, pp. 1018-1022, doi: 10.1016/j.jinorgbio.2005.01.005.

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Title A non-radioactive method for measuring Cu uptake in HepG2 cells
Author(s) Fosset, C.
McGaw, B. A.
Reid, M. D.
McArdle, H. J.
Journal name Journal of inorganic biochemistry
Volume number 99
Issue number 5
Start page 1018
End page 1022
Publisher Elsevier
Place of publication Amsterdam, The Netherlands
Publication date 2005-05
ISSN 0162-0134
Summary At present, all data on Cu uptake and metabolism have been derived from radioactive uptake experiments. These experiments are limited by the availability of the radioactive isotopes 64Cu or 67Cu, and their short half-life (12.5 and 62 h, respectively). In this paper, we investigate an alternative method to study the uptake of Cu with natural isotopes in HepG2 cells, a liver cell line used extensively to study Cu metabolism. In nature, Cu occurs as two stable isotopes, 63Cu and 65Cu (63Cu/65Cu = 2.23). This ratio can be measured accurately using inductively coupled plasma mass spectrometry (ICP-MS). In initial experiments, we attempted to measure the time course of Cu uptake using 65Cu. The change in the 63Cu/65Cu ratio, however, was too small to allow measurement of Cu uptake by the cells. To overcome this difficulty, the natural 63Cu/65Cu ratio in HepG2 cells was altered using long-term incubation with 63Cu. This had a significant effect on Cu concentration in HepG2 cells, changing it from 81.9 ± 9.46 pmol μg DNA−1 (week 1) to 155 ± 8.63 pmol μg DNA−1 (week 2) and stabilising at 171 ± 4.82 pmol μg DNA−1 (week 3). After three weeks of culture with 2 μM 63Cu the 63Cu/65Cu changed from 2.18 ± 0.05 to 15.3 ± 1.01. Cu uptake was then investigated as before using 65Cu. Uptake was linear over 60 min, temperature dependent and consistent with previous kinetics data. These observations suggest that stable isotope ICP-MS provides an alternative technique for the study of Cu uptake by HepG2 cells.
Language eng
DOI 10.1016/j.jinorgbio.2005.01.005
Field of Research 060104 Cell Metabolism
HERDC Research category C1.1 Refereed article in a scholarly journal
Persistent URL http://hdl.handle.net/10536/DRO/DU:30024808

Document type: Journal Article
Collection: Faculty of Science, Engineering and Built Environment
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