New Views on Lipid Dynamics: A Non-Equilibrium Model of Ligand-Receptor Interaction

Bergel'son, L. D.

doi:10.1007/978-3-642-61374-6_1

Cited by 2 publications

(1 citation statement)

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“…The resonance of choline from phospholipids found in the MDA-MB231 and SKBR3 cell lines was not detected in the corresponding membrane preparation. Indeed, the membranes are mainly comprised of phospholipids in the bilayer structure in which the polar head groups lie at the membrane surface (Bergelson, 1988). Due to strong interactions at the surface, the N-trimethyl ammonium groups may be strongly immobilised and therefore invisible to high resolution NMR.…”

Section: Resultsmentioning

confidence: 99%

Cell and membrane lipid analysis by proton magnetic resonance spectroscopy in five breast cancer cell lines

Moyec

Tatoud²,

Eugène³

et al. 1992

Br J Cancer

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Summary The lipid composition of five human breast cancer cell lines T47D, was assessed by proton magnetic resonance spectroscopy (MRS) in whole cells and membraneenriched fractions. The proportions of the three main lipid resonances in 1D spectra were different for each cell line. These resonances included mobile methyl and methylene functions from fatty acids of triglycerides and phospholipids and N-trimethyl from choline of phospholipids. T47D and Materials and methods Cell cultureFive human breast carcinoma cell lines were used: MCF-7, MDA-MB 231, T47D, SKBR3 and ZR-75-l (Engel & Young, 1978). All these cell lines were cultured in Dulbecco's modified Eagle's medium (DMEM, Gibco) supplemented with 10% foetal calf serum, as adherent cells.The cells were mass cultured in the same conditions, in 150 cm2 flasks. The cells from three flasks (107 cells) were harvested at confluency with a rubber policeman scraper and centrifuged at 1,000 r.p.m. for 10 min. For whole cell MRS experiments, the cells were washed three times with 2 ml of phosphate-buffered saline (PBS: KH2PO4 0.2 g 1-, KCI 0.2 g 1-', NaCl 8 g 1-', Na2HPO41.15 g 1-') in deuterium oxide (D20). The final pellet was suspended in PBS/D20 to obtain a final volume of 0.5 ml and placed in a 5 mm MRS tube. For cell membrane preparation, the same number of cells were washed in cool PBS and disrupted with a hypotonic Tris (tris[hydroxymethyl]aminoethane) buffer (0.01 M), followed by a freeze-thaw cycle at -20C; finally, polytron was used three times for 5 s. The resulting homogenate was centrifuged at 1,000 g for 10 min and the supernatant was centrifuged at 47,000 g for 30 min (Beckmann TL 100, rotor TL 100-2). For MRS experiments, the final pellet was suspended in PBS/D20 and 0.5 ml of the suspension was placed in a 5 mm MRS tube. The specific Na+/K+ ATPase activity was determined in the presence or absence of digitoxigenin using a coupled assay method previously described (Noel & Godfraind, 1984

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Section: Resultsmentioning

confidence: 99%