Induced Pluripotent Stem Cell-Derived Dendritic Cells Provide a Reliable In Vitro Platform for Functional Screening of Immunoregulatory Probiotics

Sep 30, 2026

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Immunoregulatory effects of probiotics are widely studied for maintaining immune balance, yet conventional in vitro screening depends on fresh donor-derived primary immune cells, causing inter-individual and temporal variability that reduces reproducibility. This study uses human iPSC-derived dendritic cells co-cultured with five probiotic strains to evaluate immunomodulatory interactions. Conditioned media from these co-cultures were applied to human CD4+ T cells to test effects on differentiation. ELISA showed cytokines were secreted at distinct concentrations depending on the probiotic strain, and several strains promoted CD4+ T-cell differentiation into regulatory T cells.

LanguageEnglish

CategoryHealthcare

FormatPDF

Pages18

Reading Time45 min

AccessFree to download

Introduction

Probiotics and gut microbiota–health links

Need for reliable standardized screening platforms

Materials and Methods

iPSC-derived dendritic cell co-culture with probiotic strains

Conditioned medium stimulation of CD4+ T cells

Results

Cytokine secretion profiles by probiotic strain

Promotion of regulatory T cell differentiation

Conclusions

iPSC-derived dendritic cells as a functional screening platform

Article Induced Pluripotent Stem Cell-Derived Dendritic Cells Provide a Reliable In Vitro Platform for Functional Screening of Immunoregulatory Probiotics Yin-Ling Chiang 1, Men-Yee Chiew 2, Sheng-Jye Lim 2, Ding-Li Chou 3,†, Huai-En Lu 2, * and Ching-Ping Tseng 1,3, * Academic Editor: Daniele Lana Received: 24 November 2025 Revised: 17 December 2025 Accepted: 25 December 2025 Published: 27 December 2025 Copyright: © 2025 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license. 1 Industrial Development Graduate Program of College of Engineering Bioscience, National Yang Ming Chiao Tung University, Hsinchu 300, Taiwan; [candyi8113@gmail.com](candyi8113@gmail.com) 2 Center for Regenerative Medicine and Cellular Therapy, National Yang Ming Chiao Tung University, Hsinchu 300, Taiwan; [chiewmy@nycu.edu.tw](chiewmy@nycu.edu.tw) (M.-Y.C.); [shengjye2022@nycu.edu.tw](shengjye2022@nycu.edu.tw) (S.-J.L.) 3 Department of Biological Science and Technology, College of Engineering Bioscience, National Yang Ming Chiao Tung University, Hsinchu 300, Taiwan; [gs12099@yahoo.com.tw](gs12099@yahoo.com.tw) * [Correspondence: helu@nycu.edu.tw](Correspondence: helu@nycu.edu.tw) (H.-E.L.); [cpts@nctu.edu.tw](cpts@nctu.edu.tw) (C.-P.T.)† Current address: Bio Race International Inc., Zhubei City 302005, Taiwan. Abstract The immunoregulatory effects of probiotics have been widely studied, particularly in maintaining immune balance. Conventional in vitro functional screening of probiotics relies on fresh donor-derived primary immune cells, which often exhibit significant inter-individual and temporal variability, limiting reproducibility and interpretation. As an alternative, human-induced pluripotent stem cell (iPSC)-derived dendritic cells were co-cultured with five probiotic strains in the current study to evaluate their immunomodulatory interactions. To assess whether cytokines produced by probiotic-stimulated dendritic cells can influence T cell differentiation, human CD4+ T cells were exposed to the conditioned medium derived from co-cultures. Enzyme-linked immunosorbent assay results demonstrated that iPSC-derived dendritic cells secreted cytokines at distinct concentrations in response to different probiotic strains, suggesting that these cells can distinguish between different microbial stimuli, and supporting their use in functional probiotic screening. Among the five strains tested, Lactiplantibacillus plantarum LPA-56, Limosilactobacillus reuteri RU-23, and Lactobacillus fermentum Fem-99 induced cytokine production levels that promoted the differentiation of the human CD4+ T cells into regulatory T cells. These findings demonstrate that iPSC-derived dendritic cells have immunomodulatory potential, are reliable for in vitro screening of probiotics, and offer a promising strategy for selecting potent immunoregulatory probiotic candidates. Keywords: induced pluripotent stem cell; dendritic cell; probiotic; co-culture; CD4+ T cell 1. Introduction Recent studies have demonstrated that the diversity of the gut microbiota, as well as fluctuations in specific microbial populations, is closely associated with human health [1] . With the implementation of the Human Microbiome Project and advances in metagenomic technologies, the link between gut microbiota dysbiosis caused by different dietary intakes and various diseases has been well-established [2], including irritable bowel syndrome and inflammatory bowel disease [3,4], as well as metabolic and cardiovascular conditions such as obesity, diabetes, and heart disease [5–7] . Emerging evidence has also linked gut microbiota dysbiosis to neurodevelopmental and neurodegenerative disorders, including autism, Alzheimer’s disease, and even cancer [8–10] . Therefore, modulation of the gut microbiota has been associated with reduced disease incid

FAQs

Why is iPSC-derived dendritic cell screening used instead of donor-derived primary immune cells?

Conventional screening using fresh donor-derived cells shows significant inter-individual and temporal variability, limiting reproducibility and interpretation. iPSC-derived dendritic cells aim to provide a more reliable and standardized in vitro platform.

How did the study test whether probiotic effects translate to T cell differentiation?

Human CD4+ T cells were exposed to conditioned medium derived from co-cultures of iPSC-derived dendritic cells with probiotic strains. Cytokine outputs and downstream differentiation toward regulatory T cells were assessed.

What did the cytokine results indicate across different probiotic strains?

ELISA results showed that iPSC-derived dendritic cells secreted cytokines at distinct concentrations in response to different probiotic strains, supporting the cells’ ability to distinguish between microbial stimuli.

Sep 30, 2026

Uploaded by Aurelia