RESUMO
Objective To establish a hanging drop 3D cell culture model of human colon cancer cell (HT29) in 48-well cell culture plate,at the same time,through the comparison of several cell viability detection methods to determine the appropriate one for this cell culture way.Methods HT29 cells of 2 375,3 164,4 218,5 625,7 500 and 10 000/well were seeded in the bottom of the 48-well culture plate to form droplets.After 2 d of inversion culture,the cell spheroids were formed and incubated in medium for another 3 d.The volume of cell spheroids were measured,and the absorbance (A) values were detected through APH assay,MTT assay,MTT assay after digestion,CCK-8 assay and CCK-8 assay after digestion.The results were compared among different methods.Results After 5 d of culture,the cell spheroids were formed perfectly at the density of 2 375-10 000/well,and the volumes were in good linear with the original cell inoculation number at the density of 2 375-7 500/well.The A values of APH assay,MTT assay after digestion and CCK-8 assay after digestion increased with the increase of cell inoculation amount;But the cell ball digestion process was complex,and the cell viability was damaged.However,the A values of MTT and CCK-8 assay increased slowly.Conclusion The method of a hanging drop 3D cell culture model in 48-well culture plate combining with APH assay to detect cell viability is economical,accurate and easy to operate.
RESUMO
Objective To establish a hanging drop 3D cell culture model of human colon cancer cell (HT29) in 48-well cell culture plate,at the same time,through the comparison of several cell viability detection methods to determine the appropriate one for this cell culture way.Methods HT29 cells of 2 375,3 164,4 218,5 625,7 500 and 10 000/well were seeded in the bottom of the 48-well culture plate to form droplets.After 2 d of inversion culture,the cell spheroids were formed and incubated in medium for another 3 d.The volume of cell spheroids were measured,and the absorbance (A) values were detected through APH assay,MTT assay,MTT assay after digestion,CCK-8 assay and CCK-8 assay after digestion.The results were compared among different methods.Results After 5 d of culture,the cell spheroids were formed perfectly at the density of 2 375-10 000/well,and the volumes were in good linear with the original cell inoculation number at the density of 2 375-7 500/well.The A values of APH assay,MTT assay after digestion and CCK-8 assay after digestion increased with the increase of cell inoculation amount;But the cell ball digestion process was complex,and the cell viability was damaged.However,the A values of MTT and CCK-8 assay increased slowly.Conclusion The method of a hanging drop 3D cell culture model in 48-well culture plate combining with APH assay to detect cell viability is economical,accurate and easy to operate.
RESUMO
Objective To study pharmacodynemics and pharmacokenitics of apixaban in rats and investigate the correlation between them.Mehtods The UPLC-MS/MS method was applied to determining the plasma concentration of apixaban and draw the concentration-time curve.Meanwhile,the extension rate of prothrombin time (PT) was determined to draw the effect-time curve.Then the relationship between concentration and effect could be evaluated.Results After iv administration of apixaban (2 mg/kg) in rats,the main pharmacokinetic parameters AUC0-∞ and T1/2z were (4 016.07 ± 1 160.46) μg·h/L and (2.95 ± 1.59) h,respectively.After ig administration of apixaban (10 mg/kg),the main pharmacokinetic parameters AUC0-∞,T1/2z,Cmax,Tmax and bioavailability were (17 973.48 ± 3145.30) μg·h/L,(1.52 ± 0.36) h,(4 949.12 ± 615.38) μg/L,(1.00 ± 0.71) h and 89.5％,respectively.Apixaban (10 mg/kg) significantly increased PT and the effect lasted about 2 h.The changes of apixaban plasma concentration and PT extension rate were synchronous.Conclusion Apixaban has the characteristics of high oral bioavailability and rapid absorption.There is a significant correlation between PT extension rate and its plasma concentration after ig administration of 10 mg/kg in rats.