Cytotoxicity Assays are widely used by researchers and by the pharmaceutical industry to investigate the toxicity of compounds on cellular systems. Cytotoxicity assays provide a rapid, sensitive and validated approach to quantify harmful-dose ranges of compounds and to analyze the biological effects of toxicity on living cellular systems. Additionally, cytotoxicity testing can be used for quality control purposes for lot release testing of raw materials or of manufactured drug products.
Medicilon can offer multiple readouts for quantifying the cytotoxicity of compounds, including colorimetric, fluorescent and luminescent assays that can be multiplexed with apoptosis or viability measurements. Our wide range of testing platforms includes the MultiTox-Glo Multiplex Cytotoxicity Assay and the ApoGlow, MTT, XTT, MTS, and LDH assays.
Agarose Overlay
MEM Elution
Direct Contact Cytotoxicity
Indirect Contact Agar Diffusion Test
MTT Assay (quantitative)
In vitro testing using primary cells can quickly screen for new chemical entities with serious toxicology consequences. Using primary cells derived from human tissues, the relative risk can be determined in multiple organs over a population of donors. The following in vitro cytotoxicity assays using the specified endpoints have been developed in our laboratory. With our ready inventory of human and cynomolgus monkey, cells scheduling is not an issue. Contact IVAL to begin designing your study today.
Cell viability (luminescence based ATP assay)
Cell growth and survival (MTT/WST-1 metabolism)
Apoptosis
Oxidative Stress
Cytotoxicity is typically defined as the loss of membrane integrity as measured by membrane-impermeable dye uptake or the release of intracellular enzymes into the extracellular media. Promega offers multiple readouts for quantifying cytotoxicity.
1.CytoTox-Glo Cytotoxicity Assay
The CytoTox-Glo Assay is the most sensitive method available for measuring cytotoxicity. The assay quantifies the extracellular activity of an intracellular protease (dead-cell protease) when the protease is released from membranecompromised cells. The amount of luminescence directly correlates with the percentage of cells undergoing cytotoxic stress.
2.CytoTox 96 Non-Radioactive Cytotoxicity Assay
The CytoTox 96 Assay is a colorimetric alternative to 51Cr release cytotoxicity assays. The CytoTox 96 Assay quantitatively measures lactate dehydrogenase (LDH),a stable cytosolic enzyme that is released upon cell lysis, in much the same way as 51Cr is released in radioactive assays.
3.CytoTox-ONE Homogeneous Membrane Integrity Assay
The CytoTox-ONE Assay is a fluorescent method to rapidly measure the activity of lactate dehydrogenase (LDH) released into the culture medium from cells with damaged cell membranes (non-viable cells). The amount of fluorescent product produced is proportional to the number of dead cells present.
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;
③ Toxic cell special function, slight damage to cell structure and function, but very serious damage to the whole 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 drug additions, MTT amount, MTT incubation time, DMSO dissolution time (oscillation time), etc.
After the cell 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.
The indicators for quality evaluation of cytotoxicity are preliminarily determined as the following seven:
1. Average OD: Reflects the average level of OD values between multiple wells. The larger the OD value, the more cells there are.
2. Standard deviation between the OD values of multiple holes (STDEV): reflects the dispersion of the OD values of the multiple holes. 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 OD value in the form of percentage, which is vivid and intuitive.
4. Survival rate (%Viability): Calculating the survival rate of cells under 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 that if each replicate is used as a separate experiment, then the same batch of replicates can be regarded as several batches of experiments, and the distribution of survival rates at this time. The larger the index, the less 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.
Complete range of Cytoxicity markers
Single and multiple parameters from 1 cellular sample
High throughput screening methods
Easy handling, less hands-on time
Reduced amount of test compound and cells with the multiple parameter test kits
Email : marketing@medicilon.com
Tel : +86 021 58591500
Tips : Above is part of cell cytotoxicity assay,cytotoxicity testing, in vitro cytotoxicity assay, in vivo cytotoxicity assay . You can also CONTACT US with any question or enquiry you may have. We will be happy to discuss your needs in detail and design an appropriate plan of action.
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