Initial or continuous coculture with umbilical cord-derived mesenchymal stromal cells facilitates in vitro expansion of human regulatory T-cell subpopulations
1Regenerative Medicine Institute (REMEDI) at CÚRAM Research Ireland Centre for Medical Devices, School of Medicine, University of Galway, Galway H19 TK33, Ireland.
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Summary
Coculturing regulatory T cells (Tregs) with human umbilical cord-derived mesenchymal stromal cells (hUC-MSCs) significantly enhances Treg expansion and maintains their suppressive function. This method improves the yield of specific Treg subpopulations, aiding in the development of cell therapies for immune-mediated diseases.
Area of Science:
- Immunology
- Cell Therapy
- Regenerative Medicine
Background:
- Ex vivo expanded regulatory T cells (Tregs) show promise for treating immune-mediated diseases.
- Current Treg expansion methods face challenges in achieving consistent yield and purity.
- Human umbilical cord-derived mesenchymal stromal cells (hUC-MSCs) are being explored to improve cell manufacturing.
Purpose of the Study:
- To evaluate the enhancement of primary human Treg and Treg subpopulation expansion using hUC-MSC coculture.
- To assess the impact of hUC-MSC coculture on Treg suppressive potency and marker expression.
- To investigate the differential effects of hUC-MSC coculture on naive and non-naive Treg subpopulations.
Main Methods:
- Primary human total Tregs and FACS-purified Treg subpopulations were cultured with or without hUC-MSCs.
- Cultures were supplemented with anti-CD3/anti-CD28, interleukin-2, and rapamycin for 14-21 days.
- Treg expansion, T effector cell (Teff) proliferation suppression, and marker expression (HLA-DR, CD39, ICOS, Foxp3, Helios) were analyzed using FACS.
Main Results:
- hUC-MSC coculture resulted in a 4-fold greater expansion of total Tregs.
- Treg suppressive potency remained equivalent between hUC-MSC-cocultured and control Tregs.
- Coculture significantly enhanced the expansion of all Treg subpopulations, with a greater effect on non-naive Tregs (Subtypes II and III).
- hUC-MSC coculture increased HLA-DR expression in all expanded Treg subpopulations and enhanced Foxp3+ and Helios+ percentages in non-naive Tregs.
Conclusions:
- Coculture with clinical-grade hUC-MSCs substantially boosts ex vivo Treg yield and preserves suppressive potency.
- hUC-MSC coculture modulates HLA-DR expression and enhances specific Treg subpopulations, particularly non-naive Tregs.
- These findings support the use of hUC-MSCs for improved Treg manufacturing for cell-based therapies.