Genetic Toxicology

Genetic toxicology studies are conducted early in the safety testing program of pharmaceuticals and impurities for the detection of toxic, mutagenic, clastogenic and morphological transforming activity in medical devices, chemicals, food additives, etc. RDDT Laboratories protocols are designed to according to international guidelines (OECD, FDA, EU etc) and in compliance with GLP regulations. RDDT Laboratories provide clients with a reliable turn-around time and on time delivery of results. RDDT Laboratories conducts its genetic toxicology studies in a tiered approach, starting with an AMES test, and progressing to the conduct of mammalian cell and in vivo assays, which are designed to assess the toxicological relevance of any earlier observations.

AMES

Regulatory Method

Bacterial reverse mutation test for identifying mutagenicity potential of test compound applying plate-incorporation and colony counting method.

  • 5 Strains of bacteria
  • Includes dose-range finding and main test (5 concentrations of test substance, with and without metabolic activation)

High Throughput Method:

High Throughput screening of mutagenicity potential of test compound applying liquid fluctuation and colorimetric technique.

  • 2 strains of bacteria, with and without metabolic activation (5 concentrations of test substance)
  • Reduced cost and compound requirements

Chromosome Aberration

In vitro genotoxicity test for identifying structural chromosomal aberrations as a potential of the test substance.

  • CHO-K1 cell line
  • Includes dose-range finding and main test (3 concentrations of test substance, with and without metabolic activation)

Rodent Micronucleus

In vivo genotoxicity test for detection of damage induced by test article to the chromosome.

  • Includes dose-range finding and main test with 3 dose levels of test substance
  • Includes dose-range finding and main test (3 concentrations of test substance, with and without metabolic activation).
  • Oral IP, SC, IM, IV dose routes available

CYTOTOXICITY

Cytotoxicity tests are cell-based, well established assays with easily accessible endpoints for preliminary information on the general/acute toxic potential of a test compound. Several assay approaches are employed to measure the ability of an agent to damage cells in vitro as an indicator for acute toxicity in vivo.

MTS Assay (mitochondrial activity)

The MTS cell proliferation assay is a colorimetric method to identify the cytotoxic potential of a test item. The assay measures the formation of a soluble formazan product which is directly proportional to the number of live cells in culture. This assay can be performed with HepG2 cells or L-929 cells.

LDH Leakage Assay (membrane integrity)

The LDH Leakage assay determines the membrane integrity/extent of LDH leakage from cells with a damaged membrane under the influence of test item. The assay is a fluorometric measure of the release of lactate dehydrogenase (LDH) from cells with a damaged membrane by conversion of resazurin into resorufin that is measured by change in absorbance. This assay can be performed with HepG2 or L-929 cells.

Multiple cell types may be used, depending on the pharmacological action, potential target, metabolism and mechanism, or end point of the agent.

Types of Cells Available:

  • Human hepatoma HepG2 cells
  • L-929 mouse fibroblasts

Tests conducted in compliance with GLP requirements according to ISO 10993-5 guideline.