Medicilon Cytotoxicity Test Service

2021-09-17

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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.

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 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 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 unique intake mechanisms;

③The cell’s particular function is toxic, causing slight damage to cell structure and function but severe 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 investigation. Because everything is unknown, several parameters are often adjusted together so that the influence of each parameter on the result can be more evident, and some detours have been taken. Later, the weight of the parameter’s impact on the development was gradually figured out, and the experimental plan was steadily 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 stabilized, part of the data after the cytotoxicity test protocol 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 superficial 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:

Average OD: Reflects the intermediate level of OD between multiple wells. The greater the OD, the greater the number of cells.
Standard deviation between the OD values of multiple wells (STDEV): reflects the dispersion of the OD values of the various wells. The larger the standard deviation, the greater the non-parallelism between the multiple holes and the greater the operating error.
Coefficient of variation (CV) between OD values: It also reflects the dispersion of OD values between various 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 a percentage, which is vivid and intuitive.
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.
The standard deviation of survival rate (%STDEV): This value reflects the distribution of survival rate at this time. If each replicate took 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.
Coefficient of variation of survival rate (%CV): This indicator reflects the dispersion of survival rates among various wells. The larger the value, the greater the non-parallelism between each row of cell wells (i.e., a series of diluted single wells).
Half cytotoxic concentration: refers to the concentration required to produce toxic effects on half of the cells. This indicator can reflect whether the experimental system is usually working and evaluating the toxicity of unknown drugs to cells.

Cytotoxicity Detection Method

Cytotoxicity testing 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 include detecting the activity of alkaline phosphatase and acid phosphatase in the supernatant, etc.