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Updated: Aug 11, 2026
In Vitro Assays to Evaluate the Migration, Invasion, and Proliferation of Immortalized Human First-trimester Trophoblast Cell Lines
Published on: March 5, 2019
Analysis of DNA double-strand breaks induced by DNA damaging agents in trophoblast organoids
Sawako Adachi1, Asuka Tanabe2, Yoshiyuki Tsukamoto3
1Department of Biochemistry & Molecular Genetics, Faculty of Medicine, Oita University, Yufu, Japan; Department of Obstetrics and Gynaecology, Faculty of Medicine, Oita University, Yufu, Japan.
Abstract:
DNA double-strand break (DSB) is one of the most genotoxic lesions, and unrepaired DSBs can lead to chromosomal aberrations and eventually cause carcinogenesis, ageing and dysfunctions of organs. Therefore, studies of DNA repairs and DNA damage responses (DDRs) corresponding to DSBs has been gathered intense interest. Meanwhile, understanding of the clinical symptoms how dysfunction and accelerated ageing of various organs caused by the effects of DNA damages has been less accomplished. To address this, we have developed a new method that we could investigate DSB formation in trophoblast organoids. Organoids are self-organized three-dimensional tissues that mimic the complex structures and functions of an organ but are not widely used in nucleic acids research field. Using trophoblast organoids, we have examined the DSB formation induced by the treatments with various DNA damaging agents by pulsed-field gel electrophoresis. Organoids and 2D cultured cells were treated with bleomycin, mitomycin C, camptothecin, etoposide, gemcitabine, and 5-fluorouracil, and the patterns of DSB formation in the organoids and conventional two-dimensional (2D) cultured cells were compared. The pattern of DSB formation induced by DNA damaging agents in organoids was different from that appeared in 2D cultures. To further enhance versatility, we have also established a mouse organoid system and have examined its validity. We succeeded to develop a new method that can detect damage-induced DSB formation in organoids. Application of this method would be used not only for DDRs in a tissue model but also for the risk assessment of genotoxicity.
Insights
Researchers developed a new method to detect DNA double-strand breaks (DSBs) in organoids, offering insights into DNA damage responses and genotoxicity risk assessment.
Area of Science:
- Molecular Biology
- Genetics
- Toxicology
Background:
- DNA double-strand breaks (DSBs) are highly genotoxic lesions linked to aging and cancer.
- Understanding DNA damage responses (DDRs) and clinical effects of DNA damage is crucial.
- Organoids are advanced 3D tissue models but underutilized in nucleic acid research.
Purpose of the Study:
- To develop and validate a novel method for investigating DSB formation in organoids.
- To compare DSB formation patterns in organoids versus traditional 2D cell cultures.
- To assess the utility of organoids for genotoxicity risk assessment.
Main Methods:
- Utilized trophoblast organoids and established a mouse organoid system.
- Treated organoids and 2D cultured cells with various DNA damaging agents (bleomycin, mitomycin C, etc.).
- Analyzed DSB formation using pulsed-field gel electrophoresis.
Main Results:
- Successfully developed a new method to detect damage-induced DSB formation in organoids.
- Observed distinct patterns of DSB formation in organoids compared to 2D cultures.
- Validated the method using both human trophoblast and mouse organoid systems.
Conclusions:
- The novel organoid-based method enables the study of DSB formation and DDRs in a more complex tissue model.
- This approach offers a valuable tool for genotoxicity risk assessment.
- Organoids provide a distinct platform for studying DNA damage compared to conventional 2D cell cultures.
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