Protocol for scalable production of orally deliverable R-spondin1-loaded small extracellular vesicles for WNT-signaling activation and tissue regeneration.

Authors

Lingyan Yang, Ting Jiang, Shixiang Wang, Yue Yang, Zhiping Qiao, Yun-Shen Chan

Published in

STAR protocols. Volume 7. Issue 4. Pages 104891. Oct 08, 2026. Epub Oct 08, 2026.

Abstract

R-spondin1 plays a pivotal role in potentiating canonical WNT/β-catenin signaling, a pathway critical for maintaining stem cell homeostasis and driving tissue regeneration. Here, we present a scalable, serum-free protocol for producing R-spondin1-loaded small extracellular vesicles (evRSPO1) from engineered HEK293 cells, with multimodal characterization of vesicle quality and bioactivity. We also describe steps for in vitro functional validation of the vesicles using mouse intestinal organoids and for in vivo therapeutic applications in mouse intestinal regeneration via oral delivery. For complete details on the use and execution of this protocol, please refer to Yang et al.^1,2.

PMID:
42848513
Bibliographic data and abstract were imported from PubMed on 09 Oct 2026.

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