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Iron transport in cancer cell culture suspensions measured by cell magnetophoresis.

ABSTRACT: Cell motion in a magnetic field reveals the presence of intracellular paramagnetic elements, such as iron or manganese. Under controlled field and liquid media composition, such motion previously allowed us to compare the paramagnetic contribution to cell magnetic susceptibility in erythrocytes differing in the spin state of heme associated with hemoglobin. The method is now tested on cells with less obvious paramagnetic properties: cell cultures derived from human cancers to determine if the magnetophoretic mobility (MM) measurement is sufficiently sensitive to the dysregulation of the intracellular iron metabolism as suggested by reports on loss of iron homeostasis in cancer. The cell lines included hepatocellular carcinoma (Hep 3B 2.1-7 and Hep G2), promyelocytic (HL-60) and chronic myelogenous (K-562) leukemias, histiocytic lymphoma (U-937), tongue (CAL 27) and pharyngeal (Detroit 562) carcinomas, and epitheloid carcinoma (HeLa), whose MM was measured in complete media with standard and elevated soluble iron (ferric nitrate and ferric ammonium citrate), against oxy- and met-hemoglobin erythrocytes used as controls. Different cell lines responded differently to the magnetic field and the soluble iron concentrations in culture media establishing the possibility of single cell elemental analysis by magnetophoresis and magnetic cell separation based upon differences in intracellular iron concentration.

SUBMITTER: Jin X

PROVIDER: S-EPMC3556788 | biostudies-literature | 2012 May

REPOSITORIES: biostudies-literature

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Iron transport in cancer cell culture suspensions measured by cell magnetophoresis.

Jin Xiaoxia X Chalmers Jeffrey J JJ Zborowski Maciej M

Analytical chemistry 20120426 10

Cell motion in a magnetic field reveals the presence of intracellular paramagnetic elements, such as iron or manganese. Under controlled field and liquid media composition, such motion previously allowed us to compare the paramagnetic contribution to cell magnetic susceptibility in erythrocytes differing in the spin state of heme associated with hemoglobin. The method is now tested on cells with less obvious paramagnetic properties: cell cultures derived from human cancers to determine if the ma ...[more]

PMID: 22500468

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Project description:Robust and reproducible in vitro models are required for investigating the pathways involved in fluid homeostasis in the human alveolar epithelium. We performed functional and phenotypic characterisation of ion transport in the human pulmonary epithelial cell lines NCI-H441 and A549 to determine their similarity to primary human alveolar type II cells. NCI-H441 cells exhibited high expression of junctional proteins ZO-1, and E-cadherin, seal-forming claudin-3, -4, -5 and Na+-K+-ATPase while A549 cells exhibited high expression of pore-forming claudin-2. Consistent with this phenotype NCI-H441, but not A549, cells formed a functional barrier with active ion transport characterised by higher electrical resistance (529 ± 178 ? cm2 vs 28 ± 4 ? cm2), lower paracellular permeability ((176 ± 42) ×10-8 cm/s vs (738 ± 190) ×10-8 cm/s) and higher transepithelial potential difference (11.9 ± 4 mV vs 0 mV). Phenotypic and functional properties of NCI-H441 cells were tuned by varying cell seeding density and supplement concentrations. The cells formed a polarised monolayer typical of in vivo epithelium at seeding densities of 100,000 cells per 12-well insert while higher densities resulted in multiple cell layers. Dexamethasone and insulin-transferrin-selenium supplements were required for the development of high levels of electrical resistance, potential difference and expression of claudin-3 and Na+-K+-ATPase. Treatment of NCI-H441 cells with inhibitors and agonists of sodium and chloride channels indicated sodium absorption through ENaC under baseline and forskolin-stimulated conditions. Chloride transport was not sensitive to inhibitors of the cystic fibrosis transmembrane conductance regulator (CFTR) under either condition. Channels inhibited by 5-nitro-1-(3-phenylpropylamino) benzoic acid (NPPB) contributed to chloride secretion following forskolin stimulation, but not at baseline. These data precisely define experimental conditions for the application of NCI-H441 cells as a model for investigating ion and water transport in the human alveolar epithelium and also identify the pathways of sodium and chloride transport.

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Iron transport in cancer cell culture suspensions measured by cell magnetophoresis.

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Iron transport in cancer cell culture suspensions measured by cell magnetophoresis.

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