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Updated: Aug 6, 2026
Kupffer Cell Isolation for Nanoparticle Toxicity Testing
Published on: August 18, 2015
Protocol for dynamic high-throughput cell death screening of primary phagocytes following microplastic and
Tim Leonardus Philip Skrabanja1, Joëlle Anna Zoetje Klazen1, Sarah Sayed1
1Department of Respiratory Medicine, Center of Translational Immunology, University Medical Center Utrecht, Lundlaan 6, Mail Room KC02.085.2, 3584 EA Utrecht, the Netherlands.
Abstract:
Micro- and nanoplastics (MNPs) can cross epithelial barriers of the lung and/or intestine into the bloodstream. In the body, phagocytes will be exposed to plastic particles, but they are incapable of degrading them. Here, we present a protocol for high-throughput cell death screening of primary phagocytes following MNP exposure. We describe steps for isolating primary phagocytes, plating these with MNPs, and time-lapse imaging. Further, we explain detailed procedures for image analysis using the IncuCyte S3 live-cell imaging system and analysis software. For complete details on the use and execution of this protocol, please refer to Giustarini et al.1.
Insights
Micro- and nanoplastics (MNPs) enter the bloodstream and expose phagocytes. This protocol enables high-throughput screening of phagocyte cell death after MNP exposure, aiding toxicity assessments.
Area of Science:
- Environmental Science
- Toxicology
- Cell Biology
Background:
- Micro- and nanoplastics (MNPs) pose environmental and health risks.
- MNPs can penetrate biological barriers, reaching systemic circulation.
- Phagocytes encounter MNPs but cannot degrade them, raising concerns about cellular toxicity.
Purpose of the Study:
- To develop a high-throughput protocol for assessing primary phagocyte cell death induced by MNPs.
- To provide a standardized method for evaluating the cytotoxic effects of MNPs on immune cells.
Main Methods:
- Isolation of primary phagocytes from biological samples.
- Co-culture of phagocytes with MNPs under controlled conditions.
- Time-lapse live-cell imaging using the IncuCyte S3 system for monitoring cell viability.
- Detailed image analysis procedures for quantifying cell death.
Main Results:
- The protocol allows for efficient and reproducible screening of MNP-induced phagocyte toxicity.
- Quantitative data on cell death rates can be obtained rapidly.
- The method facilitates comparative toxicity assessments of different MNP types and sizes.
Conclusions:
- This protocol offers a robust platform for investigating the impact of MNPs on phagocyte health.
- It supports the assessment of MNP-related health risks and informs regulatory strategies.
- Further research can utilize this method to explore mechanisms of MNP toxicity in immune cells.
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