Related Experiment Video

Updated: Jan 24, 2026

An Automated Method to Perform The In Vitro Micronucleus Assay using Multispectral Imaging Flow Cytometry
12:56

An Automated Method to Perform The In Vitro Micronucleus Assay using Multispectral Imaging Flow Cytometry

Published on: May 13, 2019

23.2K

The application of an in vitro micronucleus test in mouse fibroblast L929 cells

Ewa Drozd1, Irena Bubko1, Karolina Jaworska2

  • 1Department of Biochemistry and Biopharmaceuticals, National Medicines Institute, Chełmska 30/34 Str., 00-725, Warsaw, Poland.

Mutation Research. Genetic Toxicology and Environmental Mutagenesis
|May 30, 2019
PubMed

Insights

This study optimized the in vitro micronucleus assay for mouse L929 cells, establishing a reliable protocol for genotoxicity testing. The developed method enhances the scope of the cytokinesis-block micronucleus test for chemical safety assessments.

Area of Science:

  • Toxicology
  • Genetics
  • Cell Biology

Background:

  • The in vitro micronucleus assay is crucial for assessing chemical genotoxicity.
  • A standardized protocol for mouse fibroblast L929 cells was lacking.

Purpose of the Study:

  • To optimize and validate the cytokinesis-block micronucleus (CBMN) test for L929 cells.
  • To establish a reliable protocol for genotoxicity studies in this cell line.

Main Methods:

  • Optimization of cytochalasin B concentration and positive controls (MMS, colchicine, CP).
  • Evaluation of phenobarbital/5,6-benzoflavone as an S9 inducer.
  • Testing of three exposure regimens and assessment of reproducibility.
  • Identification of optimal staining methods (Giemsa, acridine orange).

Main Results:

  • 1 μg/mL cytochalasin B and specific concentrations of MMS, colchicine, and CP were effective.
  • Phenobarbital/5,6-benzoflavone proved superior to Aroclor 1254 as an S9 inducer.
  • The developed protocol demonstrated reliable results and reproducibility.

Conclusions:

  • A validated CBMN assay protocol for L929 cells was established.
  • This protocol is suitable for genotoxicity testing according to OECD Guideline #487.
  • The optimized assay expands the utility of micronucleus testing in chemical safety evaluations.
Keywords:
Cytochalasin BCytokinesis blockMicronuclei assayMouse fibroblasts L929Phenobarbital/5,6-benzoflavone

More Related Videos

Author Spotlight: Cryopreservation of Whole Blood Samples for High Throughput Analysis06:14

Author Spotlight: Cryopreservation of Whole Blood Samples for High Throughput Analysis

Published on: February 23, 2024

4.9K
Advanced 3D Liver Models for In vitro Genotoxicity Testing Following Long-Term Nanomaterial Exposure08:25

Advanced 3D Liver Models for In vitro Genotoxicity Testing Following Long-Term Nanomaterial Exposure

Published on: June 5, 2020

7.2K

Related Concept Videos

Introduction to Fibroblasts01:09

Introduction to Fibroblasts

Rudolph Virchow discovered spindle-shaped cells called fibroblasts in 1858. Inactive fibroblasts, called fibrocytes, become activated by various stimuli, such as growth factors and inflammatory cytokines. Activated fibroblasts play a crucial role in wound healing, inflammation, formation of new blood vessels, and cancer progression. Uncontrolled activation of fibroblasts results in fibrosis, the excess deposition of fibrous tissue, which can lead to scarring and affect normal organs. This...
3.9K
Clinical Applications of Epidermal Stem Cells01:19

Clinical Applications of Epidermal Stem Cells

Epidermal stem cells (EpiSCs) are mainly located at the basal layer of the epidermis. These cells repair minor injuries of the skin and replace dead skin cells. However, EpiSCs’ cannot heal severe wounds such as major burns or those from diabetes or hereditary disorders. In such cases, culturing the epidermal stem cells from the patient is possible and has yielded successful treatment options, such as laboratory-grown skin grafts. These grafts are synthesized using a patient’s own...
3.3K
In-vitro Mutagenesis01:16

In-vitro Mutagenesis

To learn more about the function of a gene, researchers can observe what happens when the gene is inactivated or “knocked out,” by creating genetically engineered knockout animals. Knockout mice have been particularly useful as models for human diseases such as cancer, Parkinson’s disease, and diabetes.
16.1K
Photoluminescence: Applications01:14

Photoluminescence: Applications

Photoluminescence offers a wide range of applications due to its inherent sensitivity and selectivity. This technique allows for both direct and indirect analyses of the analyte. Direct quantitative analysis is possible when the analyte exhibits a favorable quantum yield for fluorescence or phosphorescence. However, an indirect analysis may be feasible if the analyte is not fluorescent or phosphorescent, or if the quantum yield is unfavorable. Indirect methods include reacting the analyte with...
1.0K
Radiation: Applications01:17

Radiation: Applications

The average temperature of Earth is the subject of much current discussion. Earth is in radiative contact with both the Sun and dark space; it receives almost all its energy from the radiation of the Sun and reflects some of it into outer space. Dark space is very cold, about 3 K, so Earth radiates energy into it. For instance, heat transfer occurs from soil and grasses, the rate of which can be so rapid that frost can occur on clear summer evenings, even in warm latitudes.
The average...
1.7K
Applications of Stress01:04

Applications of Stress

Consider a structure made of a boom and a rod designed to support a load. These two components are connected by a pin and stabilized by brackets and pins. The boom and the rod are detached from their supports to assess the different stresses imposed on this structure, and a free-body diagram is drawn. Then, all the forces applied, including the load acting on the structure, are identified. The reaction forces exerted on both the boom and the rod are computed using the equilibrium equations.
The...
657