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Initial or continuous coculture with umbilical cord-derived mesenchymal stromal cells facilitates in vitro expansion of human regulatory T-cell subpopulations

Title: Initial or continuous coculture with umbilical cord-derived mesenchymal stromal cells facilitates in vitro expansion of human regulatory T-cell subpopulations
Authors: Ou, Qifeng; Cormican, Sarah; Power, Rachael; Hontz, Sarah; Hanley, Shirley A; Islam, Md Nahidul; Shaw, Georgina; Deedigan, Laura M; Horan, Emma; Elliman, Stephen J; Fazekas, Barbara; Krawczyk, Janusz; Negi, Neema; Griffin, Matthew D
Contributors: China Scholarship Council PhD Fellowship; Irish Clinical Academic Training (ICAT) Programme; Wellcome Trust; Health Research Board; Health Service Executive National Doctors Training and Planning; Health and Social Care Research and Development Division; Health Service Executive of Ireland; Irish Research Council Enterprise Partnership Postdoctoral Fellowship; European Commission; Horizon 2020 Collaborative Health Project NEPHSTROM; MedTrain; Science Foundation Ireland; European Union’s Horizon 2020 Marie Skłodowska-Curie; Science Foundation Ireland Research Centres; CÚRAM
Source: Stem Cells Translational Medicine ; volume 14, issue 6 ; ISSN 2157-6564 2157-6580
Publisher Information: Oxford University Press (OUP)
Publication Year: 2025
Description: Clinical trials have demonstrated the safety and potential efficacy of ex vivo expanded regulatory T cells (Tregs) for immune-mediated diseases. Nonetheless, achieving consistent and timely Treg yield and purity remains challenging. We aimed to evaluate the potential to enhance culture expansion of primary human total Treg (CD4+/CD25+/CD127lo) and Treg subpopulations through coculture with human umbilical cord-derived mesenchymal stromal cells (hUC-MSCs). In 14- to 21-day anti-CD3/anti-CD28-, interleukin-2-, and rapamycin-containing cultures, fluorescence-activated cell sorting (FACS)-purified total Treg underwent 4-fold greater expansion following hUC-MSC coculture. Potency to suppress T effector cell (Teff) proliferation was equivalent for hUC-MSC-cocultured and control Tregs and correlated with the expression of HLA-DR, CD39, and inducible costimulator (ICOS). The impact of hUC-MSC coculture on ex vivo expansion of 3 FACS-purified Treg subpopulations [CD45RA+ (Subtype I), CD45RA−HLA-DR+ (Subtype II), and CD45RA−HLA-DR− (Subtype III)] was then investigated. Both initial and continuous hUC-MSC coculture yielded significantly higher fold expansion of each Treg subpopulation compared to control. However, the magnitude of enhancement was substantially greater for non-naive (Subtypes II and III) than for naive (Subtype I) Treg. Coculture with hUC-MSC increased HLA-DR expression of all 3 expanded Treg subpopulations while maintaining comparable Teff suppressive potency. For non-naive Treg (Subtypes II and III), both initial and continuous hUC-MSC coculture also increased the final %Foxp3+ and %Helios+. Thus, coculture with clinical-grade hUC-MSC substantially enhances the ex vivo yield, preserves the suppressive potency, and modulates HLA-DR expression of FACS-purified Treg subpopulations with greatest effect on non-naive (CD45RA−) Treg. The findings have potential to facilitate identification, functional characterization, and manufacturing of Treg subpopulations with distinct therapeutic benefits.
Document Type: article in journal/newspaper
Language: English
DOI: 10.1093/stcltm/szaf012
Availability: https://doi.org/10.1093/stcltm/szaf012; https://academic.oup.com/stcltm/article-pdf/14/6/szaf012/63490672/szaf012.pdf
Rights: https://creativecommons.org/licenses/by/4.0/
Accession Number: edsbas.385FD6C1
Database: BASE