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Medicilon Cytotoxicity Test Service

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The Biology Department of Shanghai Medicilon has extensive experience in the field of in vitro biology, through enzyme level determination, cell level determination, cell biology, biochemistry, in vitro isotope determination, stable cell line establishment, gene knockout, RNAi and MicroRNA Technology, etc., to provide a complete set of biological services.

Cytotoxicity test
Cytotoxicity test

Medicilon Cell Level Measurement Service Project

Cytotoxicity analysis
Apoptosis detection and analysis
Cell migration analysis
Cell invasion analysis
HTRF cell level detection and analysis
Immunostaining analysis
Immunofluorescence analysis (96/384 well plate)
Reporter gene analysis (green fluorescent protein/luciferase)
Radioligand receptor binding test
Cell uptake analysis
Gene knockout analysis
MicroRNA overexpression analysis
MicroRNA knockout analysis
Application analysis of adenovirus/retrovirus/lentivirus vector
Medicilon has established more than 100 cell lines for cell-level determination.
Breast cancer cell
Colorectal cancer
Leukemia cells
liver cancer cells
Renal cancer cells
Lung cancer cell
Melanoma cells
Ovarian cancer cells
Pancreatic cells
Gastric cancer

Learn more about cytotoxicity services

Cytotoxicity refers to chemical substances (drugs) acting on the basic structure and/or physiological processes of cells, such as cell membrane or cytoskeleton structure, cell metabolism process, synthesis, degradation or release of cell components or products, ion regulation and cell division processes , Leading to disorders of cell survival, proliferation and/or function, causing adverse reactions.

Cytotoxicity Can be Divided into 3 Types According to the Mechanism of Action:

①Basic cytotoxicity, involving one or more of the above structural or functional changes, acting on all types of cells;
②Selective cytotoxicity, which exists on some differentiated cells, is mainly triggered by the biotransformation of chemical substances, binding with special receptors or special intake mechanisms;
③The cell’s special function is toxic, causing slight damage to cell structure and function, but very serious damage to the entire body.

The establishment of the cytotoxicity experiment protocol After determining the MTT method as the method of cytotoxicity experiment, we explored and optimized the various parameters that may be involved in the experiment. Initially, because everything is unknown, several parameters are often adjusted together, so that the influence of each parameter on the result is not very clear, and some detours have been taken. In the later stage, the weight of the parameter’s influence on the result was gradually figured out, and the experimental plan was gradually established.
The experimental parameters considered are: cell batch, cell density, incubation time, number of dosing, MTT amount, MTT incubation time, DMSO dissolution time (oscillation time), etc.

After the cytotoxicity test method is stabilized, part of the data after the cytotoxicity test protocol is stabilized, the standard deviation of the absorbance of each well, the coefficient of variation and the standard deviation of the inhibition rate of each well can be controlled at a very low level, indicating that the experimental protocol is stable , With good reproducibility.

Cytotoxicity Test Quality Evaluation Index

The indicators for quality evaluation of cytotoxicity are preliminarily determined as the following seven:

  1. Average OD: Reflects the average level of OD between multiple wells. The greater the OD, the greater the number of cells.

  2. Standard deviation between the OD values of multiple wells (STDEV): reflects the dispersion of the OD values of the multiple wells. The larger the standard deviation, the greater the non-parallelism between the multiple holes and the greater the operating error.

  3. Coefficient of variation (CV) between OD values: It is also an indicator reflecting the dispersion of OD values between multiple wells. Since the unit of the standard deviation (STDEV) between the OD values is the same as the unit of the OD, it cannot reflect the discrete weight, so the coefficient of variation (CV) is introduced. This indicator expresses the dispersion of the OD value in the form of percentage, which is vivid and intuitive.

  4. Survival rate (%Viability): Calculating the survival rate of cells at each drug concentration can reflect the survival of cells. The larger the value, the smaller the cytotoxic effect of the drug.

  5. Standard deviation of survival rate (%STDEV): This value reflects the distribution of survival rate at this time if each replicate is taken as a separate experiment, then the same batch of replicates can be regarded as several batches of experiments. The larger the index, the more non-parallel between the multiple holes.

  6. Coefficient of variation of survival rate (%CV): This indicator reflects the dispersion of survival rates among multiple wells. The larger the value, the greater the non-parallelism between each row of cell wells (ie a series of diluted single wells) .

  7. Half cytotoxic concentration: refers to the concentration required to produce toxic effects on half of the cells. This indicator can be used to reflect whether the experimental system is working normally and to evaluate the toxicity of unknown drugs to cells.

Cytotoxicity Detection Method

Cytotoxicity testing is mainly based on changes in cell membrane permeability. The following methods are commonly used:
MTT, XTT method: using the activity of the enzymes inside the mitochondria, specific tetrazolium salts can be transformed, and then detected by a microplate reader
LDH method: To detect cytotoxicity by detecting the enzyme activity of LDH in the cell culture supernatant
Other enzyme methods: such as detecting the activity of alkaline phosphatase and acid phosphatase in the supernatant, etc.

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