I'm performing a cytotoxicity test (LDH detection) after 24 and 48h of cell incubation on a scaffold. I assume that after 24h cell proliferation results in higher LDH content and release from cells. Can it be somehow subtracted? or maybe a relation between proliferation assay and cytotoxicity assay to determine only the influence of the substance in long period without a disturbance of higher cell number. The comparison to control group is difficult because cells have different adhesion and proliferation ratio on polystyrene well than on the scaffold.
Dear Katarzyna,
we had a similar problem. Our solution is to seed cells also on a commercially available standard scaffold as control. Results will be calculated in relation to that control scaffold (e.g. test scaffold 1 performs better/worse than control scaffold).
If you just want to assess the cytotoxicity of your scaffold you might incubate your cells with medium preconditioned with the scaffold. If any cytotoxic substances had been released the number of cells should drop. You can use also transwell approaches. In the upper chamber you put the scaffold in the lower chamber you will seed your cells. At different points in times you can analyse cell death or morphological changes of the cells. However, those approaches do not consider certain surface characteristics of your test scaffolds.
Best regards
Dirk
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