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Background <p>Microphysiological systems (MPS) are powerful tools for modeling human organ function and evaluating therapeutic interventions. The performance and translational relevance of MPS are highly dependent on the quality and suitability of the cells used. Given the expanding array of primary and stem cell-derived sources, there is a critical need for systematic frameworks to guide cell selection, particularly for disease-specific applications such as Metabolic Dysfunction-Associated Steatotic Liver Disease (MASLD).</p> Methods <p>We developed a phased, stepwise methodology to evaluate and select primary human hepatocytes for use in a liver MPS model of MASLD. The protocol incorporates assessments of cell source quality, plating efficiency, viability, baseline liver function, and responsiveness to disease-inducing conditions. Quantitative metrics and pass/fail criteria were applied at each stage to ensure consistent and reproducible evaluation across cell lots.</p> Results <p>Application of the protocol enabled effective triaging of hepatocyte sources, distinguishing cell lots with superior plating performance and functional profiles. Selected hepatocytes exhibited robust expression of liver-specific markers, maintained metabolic activity, and demonstrated disease-relevant phenotypes under MASLD-inducing conditions.</p> Discussion <p>This structured evaluation framework facilitates the identification of high-quality hepatocytes for MPS liver models, improving reproducibility and disease modeling accuracy. While this protocol was tailored for MASLD, the approach is adaptable to other liver diseases or applications. Limitations include potential variability in donor tissue availability and the need for standardization across laboratories.</p> Clinical trial number <p>Not applicable.</p>

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Road map for primary hepatocyte qualification in human liver organ models

  • 期刊论文

  • 出版模式：

    开放获取

  • 发表日期：

    2026年2月9日

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  • Mahboubeh Varmazyad,
  • Dillon C. Gavlock,
  • Michael W. Castiglione,
  • Richard DeBiasio,
  • Gregory LaRocca,
  • Celeste Reese,
  • Lawrence A. Vernetti,
  • Mark Schurdak,
  • Andrew M. Stern,
  • D. Lansing Taylor,
  • Mark T. Miedel,
  • Jacquelyn A. Brown

摘要

Background

Microphysiological systems (MPS) are powerful tools for modeling human organ function and evaluating therapeutic interventions. The performance and translational relevance of MPS are highly dependent on the quality and suitability of the cells used. Given the expanding array of primary and stem cell-derived sources, there is a critical need for systematic frameworks to guide cell selection, particularly for disease-specific applications such as Metabolic Dysfunction-Associated Steatotic Liver Disease (MASLD).

Methods

We developed a phased, stepwise methodology to evaluate and select primary human hepatocytes for use in a liver MPS model of MASLD. The protocol incorporates assessments of cell source quality, plating efficiency, viability, baseline liver function, and responsiveness to disease-inducing conditions. Quantitative metrics and pass/fail criteria were applied at each stage to ensure consistent and reproducible evaluation across cell lots.

Results

Application of the protocol enabled effective triaging of hepatocyte sources, distinguishing cell lots with superior plating performance and functional profiles. Selected hepatocytes exhibited robust expression of liver-specific markers, maintained metabolic activity, and demonstrated disease-relevant phenotypes under MASLD-inducing conditions.

Discussion

This structured evaluation framework facilitates the identification of high-quality hepatocytes for MPS liver models, improving reproducibility and disease modeling accuracy. While this protocol was tailored for MASLD, the approach is adaptable to other liver diseases or applications. Limitations include potential variability in donor tissue availability and the need for standardization across laboratories.

Clinical trial number

Not applicable.