Enhanced Utilization of Induced Pluripotent Stem Cell-Derived Human Intestinal Organoids Using Microengineered Chips

Area of Science:

• * Stem cell biology and regenerative medicine.
• * Microengineering and organ-on-a-chip technology.
• * Gastrointestinal physiology and disease modeling.

Background:

• * Induced pluripotent stem cell-derived human intestinal organoids offer potential for studying intestinal epithelium.
• * Direct assay of these complex organoid structures is challenging.
• * This study aimed to develop a more accessible platform using microengineered chips.

Purpose of the Study:

• * To incorporate epithelial cells from human intestinal organoids into a microengineered Chip.
• * To determine if these cells in the Chip were polarized and possessed major intestinal epithelial subtypes.
• * To assess the biological responsiveness of the Chip to exogenous stimuli.

Main Methods:

• * Epithelial cells were isolated from human intestinal organoids and seeded into a microengineered Chip.
• * Immunocytochemistry and in situ hybridization confirmed cell polarization and subtype presence.
• * Western blot, qPCR, and permeability assays assessed biological responses to cytokines like IFN-γ and TNF-α.

Main Results:

• * Optimal cell seeding density and flow rate for the Chip were established.
• * The Intestine-Chip exhibited polarized intestinal folds containing all major epithelial cell types, including stem cells.
• * The Chip demonstrated biological responsiveness, with cytokine exposure altering gene expression and increasing intestinal permeability.

Conclusions:

• * The developed Intestine-Chip is a polarized, multi-subtype intestinal model.
• * The platform is biologically responsive to external stimuli, validating its utility.
• * This microengineered system provides a more amenable approach to organoid technology for personalized medicine and gastrointestinal research.