Non-invasive in vivo monitoring of transplanted human intestinal organoids using bioluminescence.

Authors

Chioma Moneme, Antonio Vinicios Alves da Silva, Prisca C Obidike, Bryan A Hogg, Georgia B Brousseau, Casandra Robinson, Christian Roig-Laboy, José Kleybson de Sousa, Yuwen Zhang, Lily S Cheng, Sean R Moore

Published in

Surgery open science. Volume 32. Pages 8-16. Epub May 15, 2026.

Abstract

Xenotransplantation of pluripotent stem cell-derived human intestinal organoids (HIOs) into animal hosts models early human small intestinal development. However, traditional methods to assess HIO engraftment and expansion involve invasive, non-survival procedures. We developed a novel HIO platform featuring stable intestinal epithelial expression of a red-shifted luciferase, enabling sensitive, longitudinal in vivo tracking of HIO xenografts.
Human embryonic stem cell (hESC) line H9 was lentivirus-transduced with a plasmid encoding red-shifted luciferase under a ubiquitin C promoter. After positive selection, cells were differentiated into endoderm, mid/hindgut spheroids, and HIOs. HIO maturation and luciferase expression were confirmed by immunofluorescence prior to kidney capsule implantation in 10 adult immunodeficient SCID mice. We performed serial in vivo bioluminescent imaging over 12-weeks, comparing radiance across time points using the Kruskal-Wallis test. Following euthanasia, HIO grafts were explanted to assess size and tissue architecture.
Stable transduction did not compromise the genome integrity of hESCs, pluripotency, or maturation of hESCs into HIOs. In 9 of 10 mice, HIO radiance was detectable within two weeks of implantation. Mean radiance significantly increased over time (p = 0.03). Bioluminescence correlated with the mass of explanted HIOs upon euthanasia (ρ = 0.923, p < 0.001). Serendipitously, luciferase expression was uniform in H9 hESCs but restricted to the epithelium in HIOs pre- and post-implantation.
We successfully generated transplantable bioluminescent HIOs from pluripotent stem cells, setting the stage for dynamic monitoring of HIOs to further elucidate human intestinal development and injury response across the lifespan and other diverse applications.

PMID:
42210983
Bibliographic data and abstract were imported from PubMed on 19 Sep 2026.

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