Proton Magnetic Resonance Spectroscopy of Lymphocytes: An Historical Perspective

Mountford, Carolyn E.; Mackinnon, Wanda B.

doi:10.1006/immu.1994.1012

Cited by 13 publications

(11 citation statements)

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“…As previously described (Mountford & Tattersall, 1987) (Williams et al, 1988) and in activated cells (Holmes et al, 1990) as well as after the use of differentiating agents (Van Haaften-Day et al, 1988). The plasma membrane origin of the lipid signals was demonstrated by analysis of Chinese hamster ovary cell lines.…”

Section: Resultsmentioning

confidence: 87%

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: 87%

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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“…(For a review see Ref. 13). In cultured cell models, where the biological and genetic characteristics are established, it was possible to localize the origin of the lipid signal within the cell using a combination of MRS and chemical analysis of highly purified membrane.…”

Section: Al I*mentioning

confidence: 99%

Chemical Shift Imaging of Human Colorectal Tissue (Ex Vivo)

et al. 1996

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“…17,18 Although normal cells do not give rise to signals originating from lipids, it has been demonstrated that this technique can be utilized to detect narrow signals from lipid domains in the cell membranes of actively proliferating cells, such as embryonic and tumor, [19][20][21] as well as in differentiating myoblasts. 22 The signals in these cells can give precise information regarding membrane dynamics and thus serve as natural probes for detection of changes in membrane structure.…”

Section: Discussionmentioning

confidence: 99%

“…The measured FIDs were zerofilled to 64 K and Fourier transformed without apodization to the frequency domain. Resonance chemical shifts were expressed in ppm in reference to the internal standard TSP [sodium 3-trimethylsilyl (2,2,3,3-D 4 )propionate] assigned to 0 ppm. 7,9,10 A sample preparation and NMR measurement lasted about 20 min.…”

Section: Nmr Measurementsmentioning

confidence: 99%

“…[3][4][5][6][7] In view of the variations in membrane electrical properties observed in the K562 cell line as a consequence of adhesive growth of these cells onto the positively-charged polylysine surface, it was the purpose of the present study to elucidate the nature of the membrane perturbations noted. Since variations in membrane electrical properties may denote alterations in membrane structure, 1 H-NMR was used to establish if, in fact, such modifications had taken place.…”

Section: Introductionmentioning

confidence: 99%

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Forced adhesive growth of K562 leukemic cells that normally grow in suspension induces variations in membrane lipids and energy metabolism: A proton NMR study

Lamanna

Motta

Romano

et al. 1999

J. Biomed. Mater. Res.

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The mechanisms responsible for the adhesion of cells onto a material's surface and the effects that that adhesion may have on cell structure and function are fundamental questions in biomaterials research. We recently demonstrated that the erythroleukemic cell line K562, which normally grows in suspension, can be induced to grow attached to a polylysine-coated solid surface in an anchorage-dependent manner. In this study, the effects of the growth of K562 cells onto polylysine were further investigated utilizing 500 MHz 1H-NMR spectroscopy. The NMR results showed that when K562 cells are grown attached to a positively-charged polylysine surface, there are alterations in lipids and energy metabolism. In particular, there was a 31% increase in phosphatidylcholine and a 15% decrease in each of its two precursors, glycerophosphatidylcholine and choline, as well as a 20% increase in CH2 lipids and a 7% decrease in CH3 lipids in treated cells compared to the controls. These results suggest that adhesive growth can induce strong variations in membrane structure, including the membrane fluidity of K562 cells. In addition, in cells attached to polylysine there was about a 10% decrease in creatine (together with phosphocreatine), a 20% increase in gamma-glutamate, a 15% increase in beta-glutamate, and a 24% decrease in lactate. This second set of results, which is closely related to energy metabolism, indicates that not only does adhesive growth induce changes in K562 cell membrane structure, but also in the utilization of energy in these cells. The data are discussed in view of the possible role played by surface charge in affecting cell structure and function in cells that come into direct contact with charged biopolymers.

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Proton Magnetic Resonance Spectroscopy of Lymphocytes: An Historical Perspective

Cited by 13 publications

References 0 publications

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

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

Chemical Shift Imaging of Human Colorectal Tissue (Ex Vivo)

Forced adhesive growth of K562 leukemic cells that normally grow in suspension induces variations in membrane lipids and energy metabolism: A proton NMR study

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