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Updated: Jun 10, 2026
Human Pluripotent Stem Cell Based Developmental Toxicity Assays for Chemical Safety Screening and Systems Biology Data Generation
Published on: June 17, 2015
The Relative Semi-quantification of mRNA Expression as a Useful Toxicological Endpoint for the Identification of
K Bigot1, J de Lange, G Archer
1ECVAM, JRC Institute of Health and Consumer Protection, Ispra, Italy; FRAME Alternatives Laboratory, School of Biomedical Sciences, Nottingham University, UK.
Abstract:
Embryonic stem cells, which resemble the undifferentiated cells of the epiblast in a blastocyst, are able to differentiate into derivatives of the primary germ layers, including cardiomyocytes. The effects of embryotoxic/teratogenic compounds on the differentiation of cells was examined by semi-quantification of the mRNA expression using the RT-PCR protocol. Alpha and beta myosin heavy chain (alpha-MHC and beta-MHC, respectively) mRNA expression were chosen as tissue-specific markers, characteristic of early cardiac muscle development. Nine chemicals were investigated, chosen according to their in vivo embryotoxic/teratogenic potential in mice. The teratogens all-trans retinoic acid (RA), 5-fluorouracil (5-FU), hydroxyurea (HU), diphenylhydantoin (DPH) and caffeine (Caff) caused a significant reduction in MHC mRNA expression at a dose lower than that required for cytotoxicity. Saccharin (Sacc) had a similar effect, while penicillin G (PenG), isoniazid (Iso) and cytarabine (Ara-C) only showed an effect on MHC mRNA expression when the cells had a significant loss in viability. The inability to identify the strong teratogen Ara-C is related to the inappropriate target tissue.
Insights
Embryonic stem cells can model drug effects on early heart development. Several teratogens significantly reduced cardiac myosin heavy chain (MHC) mRNA expression, indicating potential embryotoxicity.
Area of Science:
- Developmental biology
- Toxicology
- Stem cell research
Background:
- Embryonic stem cells (ESCs) differentiate into various cell types, mirroring early embryonic development.
- Cardiac muscle development involves specific gene expression, such as alpha and beta myosin heavy chain (MHC) mRNA.
- Assessing the impact of embryotoxic compounds on cell differentiation is crucial for understanding developmental risks.
Purpose of the Study:
- To investigate the effects of known embryotoxic/teratogenic compounds on the differentiation of cardiomyocytes from ESCs.
- To evaluate the utility of mRNA expression of cardiac-specific markers as an indicator of compound-induced developmental toxicity.
Main Methods:
- Utilized semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR) to measure mRNA expression levels.
- Employed alpha- and beta-myosin heavy chain (MHC) mRNA as specific markers for early cardiac muscle development.
- Tested nine chemicals with varying in vivo embryotoxic/teratogenic potentials in mice using ESC differentiation models.
Main Results:
- Teratogens like all-trans retinoic acid (RA), 5-fluorouracil (5-FU), hydroxyurea (HU), diphenylhydantoin (DPH), and caffeine (Caff) significantly reduced MHC mRNA expression at non-cytotoxic doses.
- Saccharin (Sacc) demonstrated a similar inhibitory effect on MHC mRNA expression.
- Penicillin G (PenG), isoniazid (Iso), and cytarabine (Ara-C) affected MHC mRNA expression only at cytotoxic concentrations.
- The potent teratogen cytarabine (Ara-C) was not effectively identified due to potential issues with the chosen target tissue.
Conclusions:
- ESC-derived cardiomyocyte differentiation is a sensitive model for detecting the embryotoxic potential of certain chemicals, particularly those affecting early cardiac development.
- MHC mRNA expression serves as a valuable biomarker for assessing drug-induced cardiotoxicity during early development.
- The model's effectiveness may vary depending on the compound's mechanism of action and the specific differentiation pathway targeted.
