Improved microscopic observation of mammalian cells on microcarriers by fluorescent staining

Objective To develop an assay for viability detection of liver cell cultured on the Cytodex 3 in situ.Methods Two nucleic acid dyes,SYTO 13 and ethidium bromide(EB) were used to assess the viability of liver cells.Cells were stained with SYTO 13/EB.The viability of cells was determined by fluorescence microscope,and compared with MTT assay.Results Although the cell number and morphology could be clearly seen with the Cytodex 3,but cells viability could not be discerned.No cell was obviously visible inside the gelatin beads,although some cells were seen on the external surface in MTT assay.When cells were cultivated as aggregates,this FDA/EB staining technique could also be applied to such a system.High viability could be seen in large clumps of cells.Conclusion This staining technique is to be simple,quick and reliable when used in microcarrier liver cell culture.This technique has been applied to a number of microcarrier.The use of this technique with macroporous microcarriers makes it a potential visualization technique in other immobilized mammalian cell systems.