Direct contact between iPSC-derived macrophages and hepatocytes drives reciprocal acquisition of Kupffer cell identity and hepatocyte maturation

Ginhoux, F.; Lee, C. Z. W.; Tasnim, F.; Huang, X.; Sethi, R.; Song, Y.; Kozaki, T.; De Schepper, S.; Ang, N.; Low, I.; Hwang, Y. Y.; Chen, J.; Yu, H.

doi:10.1101/2025.08.26.672295

bioRxiv · 10.1101/2025.08.26.672295

Direct contact between iPSC-derived macrophages and hepatocytes drives reciprocal acquisition of Kupffer cell identity and hepatocyte maturation

Ginhoux, F.·Lee, C. Z. W.·Tasnim, F.·Huang, X.·Sethi, R.·Song, Y.·Kozaki, T.·De Schepper, S.·Ang, N.·Low, I.·Hwang, Y. Y.·Chen, J.·Yu, H.

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Abstract

Hepatic macrophages play central roles in liver homeostasis, injury, and immune-mediated hepatotoxicity through dynamic crosstalk with hepatocytes. While monocyte-derived macrophages have been widely used in vitro, they do not fully recapitulate the biology of liver-resident Kupffer cells (KCs), which are embryonically derived and maintained locally. Recent advances suggest that induced pluripotent stem cell (iPSC)-derived macrophages (iMacs) more closely resemble embryonic macrophages and may therefore serve as a relevant platform to model KC biology. Here, we developed a human iPSC-based co-culture system by combining iMacs with iPSC-derived hepatocytes (iHeps) derived from the same donor, enabling direct cell-cell interactions. We hypothesized that such interactions would both enhance hepatocyte maturation and promote KC-like differentiation of iMacs. Indeed, co-culture induced KC-like phenotypes in iMacs and improved functional maturation of iHeps, highlighting the importance of bidirectional cellular communication. Comparative analyses with iMacs cultured in hepatocyte-conditioned media revealed that direct contact provides additional signals beyond soluble factors in driving hepatic macrophage specialization. Functionally, this co-culture system demonstrated improved physiological relevance, particularly in modeling immune-mediated drug responses, as evidenced by enhanced cytokine production profiles upon exposure to a panel of test compounds. Overall, this study establishes a novel human iPSC-derived platform that captures key aspects of hepatocyte-macrophage crosstalk, providing a more physiologically relevant model to investigate liver biology and assess immune-mediated drug toxicity.

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Ginhoux, F., Lee, C. Z. W., Tasnim, F., Huang, X., Sethi, R., Song, Y., Kozaki, T., De Schepper, S., Ang, N., Low, I., Hwang, Y. Y., Chen, J., Yu, H.. 2025-08-30. Direct contact between iPSC-derived macrophages and hepatocytes drives reciprocal acquisition of Kupffer cell identity and hepatocyte maturation. https://doi.org/10.1101/2025.08.26.672295

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