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Published on: May 11, 2021
Mouse embryonic stem cell adherent cell differentiation and cytotoxicity (ACDC) assay
Marianne Barrier1, Susan Jeffay, Harriette P Nichols
1US EPA, ORD, NHEERL, ISTD, SBB, MD-72, Research Triangle Park, NC 27711, USA. Barrier.Marianne@epa.gov
Reproductive Toxicology (Elmsford, N.Y.)
|February 8, 2011
Summary
A new assay, the adherent cell differentiation and cytotoxicity (ACDC) assay, effectively evaluates chemical impacts on mouse embryonic stem cell (mESC) viability and differentiation. This method aids in distinguishing direct toxicity from effects on stem cell development.
Area of Science:
- Toxicology
- Developmental Biology
- Stem Cell Research
Background:
- Evaluating chemical effects on stem cells requires methods to assess both viability and differentiation.
- Existing methods may not efficiently distinguish between cytotoxicity and differentiation inhibition.
Purpose of the Study:
- To develop and validate a novel assay, the Adherent Cell Differentiation and Cytotoxicity (ACDC) assay.
- To assess the utility of the ACDC assay in evaluating chemical-induced effects on mouse embryonic stem cells (mESCs).
Main Methods:
- Developed the ACDC assay using pluripotent J1 mESCs cultured for 9 days.
- Employed in-cell western technique with DRAQ5/Sapphire700 stains for cell quantification.
- Used myosin heavy chain protein as a cardiomyocyte differentiation marker, corrected for cell number.
Main Results:
- The ACDC assay successfully separated cytotoxicity from differentiation effects.
- Evaluated compounds (acetic acid, 5-fluorouracil, bromochloroacetic acid) showed distinct potencies via ACDC and embryonic stem cell test.
- TaqMan low-density arrays characterized gene marker changes during differentiation and chemical exposure.
Conclusions:
- The ACDC assay is a robust technique for evaluating xenobiotic effects on mESC differentiation and cell number.
- This single assay provides a comprehensive assessment of chemical impacts on stem cell fate and viability.
