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The use of three-dimensional (3D) models, such as spheroids, offers a valuable approach to mimicking the physiological and pathological characteristics of human gastric tissues. These models are particularly relevant for studying food contamination and ingestion-related toxicity. In this protocol, Spheroids are formed in nonadherent agarose-coated 96-well plates by seeding defined cell densities, without cell adherence to the substrate, thus promoting self-aggregation. Over a 7-day culture period, spheroid diameter is monitored daily to assess compaction and stability. This method results in uniform spheroids ranging between 150 and 350 μm, suitable for downstream toxicological assays. This protocol is cost-effective, user-friendly, and adaptable to various experimental conditions.

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Advanced In Vitro Models to Assess Contaminants Toxicity by Ingestion

  • 章节

  • 发表日期：

    2026年

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  • Fernanda Rosário,
  • Helena Oliveira

摘要

The use of three-dimensional (3D) models, such as spheroids, offers a valuable approach to mimicking the physiological and pathological characteristics of human gastric tissues. These models are particularly relevant for studying food contamination and ingestion-related toxicity. In this protocol, Spheroids are formed in nonadherent agarose-coated 96-well plates by seeding defined cell densities, without cell adherence to the substrate, thus promoting self-aggregation. Over a 7-day culture period, spheroid diameter is monitored daily to assess compaction and stability. This method results in uniform spheroids ranging between 150 and 350 μm, suitable for downstream toxicological assays. This protocol is cost-effective, user-friendly, and adaptable to various experimental conditions.