Evaluation of drug and chemical toxicity with cell culture

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BACKGROUND

Human body is introduced with a vast range of drugs and chemicals every day. Toxicologists and safety professionals require quick and reliable information to identify the risk of harmful drugs and chemicals. To estimate the toxicity of drugs and chemicals, cell viability assay is an appropriate method. Assay for quantitative cytotoxicity can be performed in various methods to determine the cell viability- MTT assay, LDH assay, neutral red assay, and calcein assay.

Aim of this experiment is to evaluate the toxicity of drugs and chemicals in cell culture system with MTT assay method.

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REQUIREMENTS  

Sample:      HeLa cells, drug or chemicals (for evaluation)

Apparatus:  35 mm sterile petri dish,

Multi-Channel Pipette,

Microplate,

Spectrophotometer,

Laminar flow hood.

Chemicals:  MTT assay kit,

  HBSS

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PROCEDURE

Harvest the cells at 10³-10⁴ per well further 6 to 24 hours of incubation at 37°C. Add the drug or chemical to be tested in dose and time dependent. After that you can observe the cells in the microscope and wash with HBSS further addition of 10 μL MTT Reagent per well and incubate until purple precipitate is visible (approximately 2-4 hours) then add Detergent Reagent (100 μL). Store in the dark for 2 hours at room temperature, record absorbance at 570 nm.

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CONCLUSION

This experiment was concluded the evaluation of the drug and chemical toxicity in the HeLa cells.

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REFERENCES

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