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Updated: Aug 21, 2026
Two-dimensional Porcine Intestinal Organoids Reflecting the Physiological Properties of Native Gut
Published on: January 31, 2025
Age-dependent functional and transcriptional differences in intestinal mesenchymal stromal cells during early
Hammed Ayansola1, Younggeon Jin1
1Department of Animal and Avian Sciences, University of Maryland, College Park, MD, United States.
Insights
Early piglet intestinal mesenchymal stromal cells (iMSCs) show age-dependent functional and transcriptional shifts. Calcium channel activity in 7-day-old iMSCs influences niche factor expression, impacting epithelial development.
Area of Science:
- Gastroenterology
- Developmental Biology
- Cell Biology
Background:
- Intestinal mesenchymal stromal cells (iMSCs) are crucial for postnatal gut development.
- The temporal dynamics of iMSCs in early piglet development are not well understood.
Purpose of the Study:
- To investigate functional and transcriptional changes in jejunal iMSCs from piglets at 0, 7, and 21 days old.
- To explore the role of iMSCs in modulating epithelial development during early postnatal stages.
Main Methods:
- Enteroid-iMSC co-culture system to assess iMSC-mediated epithelial growth.
- Quantitative PCR (qPCR) and bulk RNA sequencing to analyze iMSC gene expression.
- Pharmacological inhibition of calcium channels in iMSCs.
Main Results:
- Day 0 iMSCs promoted cystic enteroids, while Day 7 iMSCs induced multi-budded structures, indicating age-dependent modulation.
- Distinct gene expression patterns were observed, with RSPO3 high at Day 0, BMP4 and PDGFRα peaking at Day 7, and antagonists increasing at Day 21.
- Day 7 iMSCs showed enrichment for calcium channel genes and related pathways; calcium channel inhibition altered niche factor expression.
Conclusions:
- iMSCs exhibit significant age-related functional and transcriptional differences during early piglet intestinal development.
- Calcium channel-related processes are implicated in the functional profile of Day 7 iMSCs.
- These findings provide a basis for understanding iMSC-epithelial interactions in early gut development.
Introduction:
Intestinal mesenchymal stromal cells (iMSCs) regulate postnatal epithelial development, yet their temporal dynamics remain poorly defined in early postnatal piglets. Here, we investigated functional and transcriptional changes in jejunal iMSCs isolated from 0-, 7-, and 21-Day-old piglets.
Methods And Results:
In an enteroid-iMSC coculture system, Day 0 iMSCs supported predominantly cystic enteroids with the highest surface area and proliferation index, whereas Day 7 iMSCs increased multi-budded enteroid morphology with intermediate proliferation, showing age-dependent differences in iMSC-mediated epithelial growth modulation. Relative real-time qPCR revealed that these functional differences aligned with niche-associated expression patterns in iMSCs. Specifically, RSPO3 level was highest at Day 0, BMP4 and PDGFRα expression peaked at Day 7, and WNT and BMP antagonists (SFRP1 and GREM1, respectively) increased in Day 21 iMSC samples. Bulk RNA sequencing further revealed shifts in iMSC transcriptomes across developmental stages, with Day 7 iMSCs exhibiting a transitional transcriptional profile enriched for genes associated with cytoskeletal reorganization and cell fate regulation compared with Day 0 iMSCs. Notably, pathway enrichment and gene set analyses identified multiple calcium channel genes and related pathways in Day 7 iMSCs compared with Day 0 iMSCs. Short-term pharmacological inhibition of calcium channels in Day 7 iMSCs was associated with rapid changes in niche factor levels, including decreased BMP4 and increased RSPO3 and WNT expressions.
Conclusion:
Together, these findings suggest age-related functional and transcriptional differences in iMSCs across early postnatal intestinal development. Our results highlight that calcium channel-related processes may be associated with changes observed in the Day 7 iMSC profile, providing a framework for future studies on iMSC-epithelial interactions during early intestinal development in piglets.
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