TY - JOUR
T1 - Intracellular volume and apparent diffusion constants of perfused cancer cell cultures, as measured by NMR
AU - Pilatus, Ulrich
AU - Shim, Hyunsuk
AU - Artemov, Dmitri
AU - Davis, Delphine
AU - Van Zijl, Peter C.M.
AU - Glickson, Jerry D.
PY - 1997/6
Y1 - 1997/6
N2 - Diffusion NMR spectroscopy was used to study intracellular volume and apparent water diffusion constants in different cell lines (DU145, human prostate cancer; AT3, rat prostate cancer; MCF-7, human breast cancer; RIF- 1, mouse fibrosacroma). The cells were grown on various matrices (collagen sponge, collagen beads, polystyrene beads) which enabled continuous growth in perfused high density cell culture suitable for NMR studies. In perfused cell systems, the attenuation of the water signal versus the squared gradient strength was fitted by the sum of two decaying exponentials. For the slowly decaying component the apparent water diffusion constant at 37°C was 0.22 (±0.02) x 10-9 s/m2 for all cell lines at diffusion times > 100 ms. It continuously increased up to 0.47(±0.05) x 10-9 s/m2 when the diffusion time was decreased to 8 ms, indicating restricted diffusion. No significant effect of the matrices was observed. The fractional volume of the slew component as determined from the biexponential diffusion curve correlated with the relative intracellular volume, as obtained from the cell density in the sample and the cell size as measured by light microsocopy. Therefore, this simple NMR approach can be used to determine intracellular volume in perfused cell cultures suitable for NMR studies. Using this information in combination with spectroscopic data, changes in intracellular metabolite concentration can be detected even when the cellular volume is changing during the experiment. The apparent diffusion constant for the fast diffusing component varied with growth matrix, cell density and cell type and also showed the typical characteristics of restricted diffusion (increase of apparent diffusion constant with time).
AB - Diffusion NMR spectroscopy was used to study intracellular volume and apparent water diffusion constants in different cell lines (DU145, human prostate cancer; AT3, rat prostate cancer; MCF-7, human breast cancer; RIF- 1, mouse fibrosacroma). The cells were grown on various matrices (collagen sponge, collagen beads, polystyrene beads) which enabled continuous growth in perfused high density cell culture suitable for NMR studies. In perfused cell systems, the attenuation of the water signal versus the squared gradient strength was fitted by the sum of two decaying exponentials. For the slowly decaying component the apparent water diffusion constant at 37°C was 0.22 (±0.02) x 10-9 s/m2 for all cell lines at diffusion times > 100 ms. It continuously increased up to 0.47(±0.05) x 10-9 s/m2 when the diffusion time was decreased to 8 ms, indicating restricted diffusion. No significant effect of the matrices was observed. The fractional volume of the slew component as determined from the biexponential diffusion curve correlated with the relative intracellular volume, as obtained from the cell density in the sample and the cell size as measured by light microsocopy. Therefore, this simple NMR approach can be used to determine intracellular volume in perfused cell cultures suitable for NMR studies. Using this information in combination with spectroscopic data, changes in intracellular metabolite concentration can be detected even when the cellular volume is changing during the experiment. The apparent diffusion constant for the fast diffusing component varied with growth matrix, cell density and cell type and also showed the typical characteristics of restricted diffusion (increase of apparent diffusion constant with time).
KW - Apparent diffusion constant
KW - Cell volume
KW - Diffusion-weighted NMR spectroscopy
KW - Microcarrier cell culture
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U2 - 10.1002/mrm.1910370605
DO - 10.1002/mrm.1910370605
M3 - Article
C2 - 9178232
AN - SCOPUS:0030960014
SN - 0740-3194
VL - 37
SP - 825
EP - 832
JO - Magnetic resonance in medicine
JF - Magnetic resonance in medicine
IS - 6
ER -