Fresh medium was added to inactivate the trypsinisation process and for detachment of cells. The suspended cells were split 1:3 every 3-4 days. Cells were grown to subconfluency and harvested as described for HepG2 cells. Kratom Denver Miami c in a water bath (1 min) in order to minimises the toxic effects exerted by the cryoprotectant DMSO on thawed cells. The content of the vial (1.
IC50 values (Inhibition concentration that caused 50% cell death) of 24 hr treatment with MSE and MIT treated cell lines. The values were interpolated from percentage dead cells curves obtained from the Trypan
blue exclusion experiments. MIT (Molar) 7. MSE and MIT. From these estimates it appears that the SH-SY5Y cells are the most sensitive of those examined to the cytotoxic and possibly cytostatic effect of MSE.
Single cultures were established for each treatment concentration and in triplicate for vehicle control. From this cell suspension preparation 4. C (5% CO2) in a shaking incubator for 3 hour (to prevent cells from settling).
Other well known assays includes MTT assay (3-(45-dimethylthiazol-2-yl)-25diphenyltetrazolium bromide) which is a metabolic assay in which tetrazolium salt is metabolised by mitochondrial dehydrogenase enzyme to form dark blue formazan in living cells. Therefore the level of colorimetric detection of formazan is proportional to the number of surviving cells (Mosman 1983). A longer term assessment for determining the capability of cells to retain the capacity for proliferating after treatment with cytotoxic agents is the clonogenicity assay.
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Various ways of cell death : Apoptosis vs necrosis Cell death represents an ultimate cycle for any kratom stores nj living organism and the equilibrium between cell
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