Characterization of a microcarrier cell culture system for 23Na MR spectroscopy studies.
A MR spectroscopy method is described for the simultaneous discrimination and observation of sodium from the three compartments created by an intact cell monolayer. Results are reported for Madin Darby Canine Kidney (MDCK) cells, an epithelial-like continuous cell line, cultured on Cytodex 1 microcarrier beads and perfused with medium containing 6 mM dysprosium (III) tripolyphosphate [Dy(TPP)2(7-)] as shift reagent. The sodium spectrum shows three resonances which are assigned to the shifted intrabead (basolateral) and extrabead (apical) pools and the unshifted intracellular pool. Ouabain inhibition of the Na(+)-K(+)-ATPase cellular pump mechanism was used to demonstrate the sensitivity of the method for monitoring intracellular sodium. The supported MDCK cells in this system remained viable after exposure for 5 h to medium containing Dy(TPP)2(7-) at a concentration of 6 mM, as determined by trypan blue dye exclusion and by comparison of the log growth rate and ability to form domes in subsequent generations of exposed cells vs unexposed controls.
Shedd
S F
SF
Department of Biomedical Engineering, Duke University, Durham, NC 27710.
Spicer
L D
LD
D000818:Animals; D002470:Cell Survival; D002478:Cells, Cultured; D004285:Dogs; D004355:Drug Stability; D004419:Dysprosium; D007202:Indicators and Reagents; D009682:Magnetic Resonance Spectroscopy / Q000379:methods*; D011122:Polyphosphates; D012964:Sodium / Q000032:analysis*
D007202:Indicators and Reagents; D011122:Polyphosphates; D004419:Dysprosium; C048543:bis(tripolyphosphate)dysprosium(III); D012964:Sodium