Protocol for a liver-bone organoid platform to study senescence-driven interorgan crosstalk

Abstract

Liver-bone crosstalk is increasingly recognized as an important regulatory axis in aging-associated diseases, such as metabolic liver disease and osteoporosis. However, conventional two-dimensional co-culture systems and animal models do not adequately capture the multicellular architecture, extracellular matrix context, and bidirectional secretory communication that underlie interorgan senescence propagation. Here, we describe a protocol for constructing a senescent liver- bone organoid platform by integrating a bone matrix-inspired three-dimensional (3D) bioprinted bone organoid system with a mouse duct-derived liver organoid culture system, followed by engineered senescence induction and conditioned-medium exchange. In this protocol, bone marrow-derived mesenchymal stem cells are incorporated into a gelatin/alginate/hydroxyapatite hybrid bioink and printed into porous 3D constructs, whereas liver ductal structures are embedded in Matrigel and expanded into liver organoids before directed differentiation. Senescence is induced in both organoid types using doxorubicin, and systemic aging-related signals are modeled by treating organoids with serum from older mice. Bidirectional communication is then interrogated through reciprocal conditioned-medium transfer assays. This platform enables investigation of interorgan senescence propagation, liver-bone axis dysfunction, and candidate mediators such as 27-hydroxycholesterol in a physiologically relevant 3D setting.

Keywords

Liver-bone organoid / Liver-bone crosstalk / Senescence / 3D bioprinting

Acknowledgments

None.

Funding

This study was funded by Noncommunicable Chronic Diseases-National Science and Technology Major Project (2024ZD0530900).

Conflict of interest

Ming He serves as an Editorial Board Member of this journal, but was not in any way involved in the editorial and peer-review process conducted for this paper, directly or indirectly. The authors declare that they have no competing interests.

Author contributions

Conceptualization: Ming He

Data curation: Yingting Zhang

Formal analysis: Yingting Zhang

Funding acquisition: Ming He

Investigation: Yingting Zhang

Project administration: Ming He

Supervision: Ming He

Writing-original draft: Jian Wang, Yingting Zhang

Writing-review & editing: Ming He

Ethics approval and consent to participate

No new human participants, human tissue, or animal experiments were performed for the preparation of this protocol manuscript. However, implementation of the protocol involves animal-derived tissues or serum and should be conducted only after approval by the relevant institutional animal care and use committee.

Consent for publication

Not applicable.

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