Optimization and Validation of FePro Cell Labeling Method 英文参考文献.docVIP

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Optimization and Validation of FePro Cell Labeling Method 英文参考文献.doc

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Optimization and Validation of FePro Cell Labeling Method 英文参考文献

OptimizationandValidationofFeProCellLabeling Method BranislavaJanic1*,AliM.Rad1,ElaineK.Jordan2,A.S.M.Iskander1,MdM.Ali1,N.RaviS.Varma1, JosephA.Frank2,3,AliS.Arbab1 1CellularandMolecularImagingLaboratory,DepartmentofRadiology,HenryFordHospital,Detroit,Michigan,UnitedStatesofAmerica,2FrankLaboratory,Radiology andImagingSciencesClinicalCenter,TheNationalInstitutesofHealth,Bethesda,Maryland,UnitedStatesofAmerica,3IntramuralResearchProgram,NationalInstituteof BiomedicalImagingandBioengineering,Bethesda,Maryland,UnitedStatesofAmerica Abstract Currentmethodtomagneticallylabelcellsusingferumoxides(Fe)-protamine(Pro)sulfate(FePro)isbasedongenerating FeProcomplexesinaserumfreemediathatarethenincubatedovernightwithcellsfortheefficientlabeling.However,this labeling technique requires long (.12–16hours) incubation time and uses relatively high dose of Pro (5–6 mg/ml) that makes large extracellular FePro complexes. These complexes can be difficult to clean with simple cell washes and may createlowsignalintensityonT2*weightedMRIthatisnotdesirable.Thepurposeofthisstudywastorevisethecurrent labeling method by using low dose of Pro and adding Fe and Pro directly to the cells before generating any FePro complexes.Humantumorglioma(U251)andhumanmonocyticleukemiacell(THP-1)lineswereusedasmodelsystemsfor attachedandsuspensioncelltypes,respectivelyanddosedependent(Fe25to100 mg/mlandPro0.75to3mg/ml)andtime dependent (2 to 48h) labeling experiments were performed. Labeling efficiency and cell viability of these cells were assessed.Prussianbluestainingrevealedthatmorethan95%ofcellswerelabeled.IntracellularironconcentrationinU251 cellsreached,30–35pg-iron/cellat24hwhenlabeledwith100mg/mlofFeand3mg/mlofPro.However,comparable labelingwasobservedafter4hacrossthedescribedFeProconcentrations.Similarly,THP-1cellsachieved,10pg-iron/cell at48hwhenlabeledwith100mg/mlofFeand3mg/mlofPro.Again,comparablelabelingwasobservedafter4hforthe describedFeProconcentrations.FeProlabelingdidnotsignificantlya