ABSTRACT
The China Food and Drug Administration (CFDA) modulated the Drug Registration Regulation at 2016.In this article,the registration classification of chemical drug was analyzed,and the definition,cormotation and technical standard of new drugs and generic drugs were compared.Compared with the 2007 version of regulations,obvious changes have happened,the scope of new drugs has been narrowed,and the definition is more strict and accurate,the scope of generic drugs has been expanded.The new regulation keeps the same evaluation standards with the ICH,FDA and EMA.Regulatory changes have a profound impact on the medical research and development:promoting the reformulation of domestic pharmaceutical market,encouraging R&D and innovation in enterprises and accelerating the pace towards the international market.
ABSTRACT
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.
ABSTRACT
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.