Status and Future of 3D Cell Culture in Toxicity Testing

Otieno, Monicah A.; Gan, Jinping; Proctor, William R.

doi:10.1007/978-1-4939-7677-5_12

Cited by 6 publications

(5 citation statements)

References 34 publications

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“…In an effort to replace, reduce and refine the reliance on animal experimentation, substantial efforts have been placed on developing 3D in vitro model systems that demonstrate greater physiological relevance and that mimic the liver-like functionality and metabolism seen in vivo . It is thought that the enhanced features of 3D models are due to the compact density of the spheroid, complex cell to cell interactions and signalling, making them an ideal candidate to improve the state-of-the-art for genotoxicological testing ( 25 ). Using liver-specific functionality assays, our data demonstrates that 3D HepG2 spheroids maintain functionality for up to 14 days in culture as fully formed spheroids.…”

Section: Discussionmentioning

confidence: 99%

Adaptation of the in vitro micronucleus assay for genotoxicity testing using 3D liver models supporting longer-term exposure durations

et al. 2020

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Following advancements in the field of genotoxicology, it has become widely accepted that 3D models are not only more physiologically relevant but also have the capacity to elucidate more complex biological processes that standard 2D monocultures are unable to. Whilst 3D liver models have been developed to evaluate the short-term genotoxicity of chemicals, the aim of this study was to develop a 3D model that could be used with the regulatory accepted in vitro micronucleus (MN) following low-dose, longer-term (5 days) exposure to engineered nanomaterials (ENMs). A comparison study was carried out between advanced models generated from two commonly used liver cell lines, namely HepaRG and HepG2, in spheroid format. While both spheroid systems displayed good liver functionality and viability over 14 days, the HepaRG spheroids lacked the capacity to actively proliferate and, therefore, were considered unsuitable for use with the MN assay. This study further demonstrated the efficacy of the in vitro 3D HepG2 model to be used for short-term (24 h) exposures to genotoxic chemicals, aflatoxin B1 (AFB1) and methyl-methanesulfonate (MMS). The 3D HepG2 liver spheroids were shown to be more sensitive to DNA damage induced by AFB1 and MMS when compared to the HepG2 2D monoculture. This 3D model was further developed to allow for longer-term (5 day) ENM exposure. Four days after seeding, HepG2 spheroids were exposed to Zinc Oxide ENM (0–2 µg/ml) for 5 days and assessed using both the cytokinesis-block MN (CBMN) version of the MN assay and the mononuclear MN assay. Following a 5-day exposure, differences in MN frequency were observed between the CBMN and mononuclear MN assay, demonstrating that DNA damage induced within the first few cell cycles is distributed across the mononucleated cell population. Together, this study demonstrates the necessity to adapt the MN assay accordingly, to allow for the accurate assessment of genotoxicity following longer-term, low-dose ENM exposure.

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Section: Discussionmentioning

confidence: 99%

Adaptation of the in vitro micronucleus assay for genotoxicity testing using 3D liver models supporting longer-term exposure durations

et al. 2020

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“…These models not only recapitulate whole organ physiology compared to 2D models, but they are also suitable for repeated exposures and chronic drug testing, an important consideration for NBM pre-clinical assessment [19]. 3D hepatic models have been shown to exhibit similar patterns of NBM transport, adsorption, and distribution that are observed in the human liver [174,175], and they also remain both viable and functional for lengthy culture periods, a characteristic which renders them useful for repeated-dose and chronic hepatotoxicity assessment of NMBs [176][177][178]. Outside of their uses in toxicity screening of NBMs, these newer models also give better insights into the pathogenesis of liver diseases such as NAFLD and steatosis, therefore unlocking the potential for shifting the in vitro NMB testing from hepatotoxicity testing to simultaneous toxicity and efficacy screening [179].…”

Section: Advanced 3d Cell Culture Models Vs Conventional Pre-clinical...mentioning

confidence: 99%

Three-dimensional (3D) liver cell models - a tool for bridging the gap between animal studies and clinical trials when screening liver accumulation and toxicity of nanobiomaterials

Tutty

Movia

Prina-Mello

2022

Drug Deliv. and Transl. Res.

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Despite the exciting properties and wide-reaching applications of nanobiomaterials (NBMs) in human health and medicine, their translation from bench to bedside is slow, with a predominant issue being liver accumulation and toxicity following systemic administration. In vitro 2D cell-based assays and in vivo testing are the most popular and widely used methods for assessing liver toxicity at pre-clinical stages; however, these fall short in predicting toxicity for NBMs. Focusing on in vitro and in vivo assessment, the accurate prediction of human-specific hepatotoxicity is still a significant challenge to researchers. This review describes the relationship between NBMs and the liver, and the methods for assessing toxicity, focusing on the limitations they bring in the assessment of NBM hepatotoxicity as one of the reasons defining the poor translation for NBMs. We will then present some of the most recent advances towards the development of more biologically relevant in vitro liver methods based on tissue-mimetic 3D cell models and how these could facilitate the translation of NBMs going forward. Finally, we also discuss the low public acceptance and limited uptake of tissue-mimetic 3D models in pre-clinical assessment, despite the demonstrated technical and ethical advantages associated with them. Graphical abstract 3D culture models for use as in vitro alternatives to traditional methods and conventional in vivo animal testing for testing liver accumulation and toxicity of nanobiomaterials

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“…These models not only recapitulate whole organ physiology, but they are also suitable for repeated exposures and chronic drug testing, an important consideration for NBM pre-clinical assessment [20]. 3D hepatic models have been shown to exhibit similar patterns of NBM transport, adsorption and distribution that are observed in the human liver [140,141], and they also remain both viable and functional for lengthy culture periods, a characteristic which renders them useful for repeated-dose and chronic hepatotoxicity assessment of NMBs [142][143][144]. Outside of their uses in toxicity screening of NBMs, these newer models also give better insights into the pathogenesis of liver diseases such as NAFLD and steatosis, therefore unlocking the potential for shifting the in vitro NMBs testing from hepatotoxicity testing to simultaneous toxicity and e cacy screening [145].…”

Section: Relevance Of In Vitro Models Vs In Vivomentioning

confidence: 99%

Three-dimensional (3D) liver cell models - A Tool for Bridging the Gap Between Animal Studies and Clinical Trials When Screening Liver Accumulation and Toxicity of Nanobiomaterials

Tutty

Movia

Prina‐Mello

2022

Preprint

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Despite the exciting properties and wide-reaching applications in human health and medicine of nanobiomaterials (NBMs), their translation from bench to bedside is slow, with a predominant issue being liver accumulation and toxicity following systemic administration. In vitro 2D cell-based assays and in vivo testing are the most popular and widely used methods for assessing liver toxicity at preclinical stages, however these fall short in predicting toxicity for NBMs. Focusing on the in vitro and in vivo assessment, the accurate prediction of human-specific hepatotoxicity is still a significant challenge to researchers. This review describes such methods, focusing on the limitations they bring in the assessment of NBM hepatotoxicity, as one of the reasons defining the poor translation for NBMs. We will then present some of the most recent advances towards the development of more biologically relevant in vitro liver methods based on tissue-mimetic 3D cell models and how these could facilitate the translation NBM going forward. Finally, we also discuss the low public acceptance and limited uptake of tissue-mimetic 3D models in the pre-clinical assessment, despite the demonstrated technical and ethical advantages.

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Status and Future of 3D Cell Culture in Toxicity Testing

Cited by 6 publications

References 34 publications

Adaptation of the in vitro micronucleus assay for genotoxicity testing using 3D liver models supporting longer-term exposure durations

Adaptation of the in vitro micronucleus assay for genotoxicity testing using 3D liver models supporting longer-term exposure durations

Three-dimensional (3D) liver cell models - a tool for bridging the gap between animal studies and clinical trials when screening liver accumulation and toxicity of nanobiomaterials

Three-dimensional (3D) liver cell models - A Tool for Bridging the Gap Between Animal Studies and Clinical Trials When Screening Liver Accumulation and Toxicity of Nanobiomaterials

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