Basic Study
Copyright ©The Author(s) 2025. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Stem Cells. Jul 26, 2025; 17(7): 104607
Published online Jul 26, 2025. doi: 10.4252/wjsc.v17.i7.104607
Differentiation of patient-specific induced pluripotent stem cells derived from type 1 diabetes peripheral blood mononuclear cells into pancreatic β-like cells
Kun Wang, Wei Lin, Jun-Yong Han, Jin-Yan Chen, Rong-Hua Liu, Zhen Yu, Jing-Jun Jin
Kun Wang, Jun-Yong Han, Jin-Yan Chen, Rong-Hua Liu, Zhen Yu, Jing-Jun Jin, Fujian Key Laboratory of Medical Analysis, Fujian Academy of Medical Sciences, Fuzhou 350001, Fujian Province, China
Wei Lin, Department of Endocrinology, Shengli Clinical College of Fujian Medical University, Fuzhou University Affiliated Provincial Hospital, Fuzhou 350001, Fujian Province, China
Author contributions: Wang K and Lin W conceived of the project and wrote the manuscript; Han JY, Chen JY, and Jin JJ also contributed to study conception and design; Lin W performed sample collection and analysis; Wang K, Han JY, Liu RH, and Yu Z performed material preparation, data analysis, and conducted most of the experiments; Chen JY and Jin JJ provided critical discussions and advice; Wang K and Han JY edited the manuscript. All authors have read and agreed to the published version of the manuscript.
Supported by the Nonprofit Research Institutes Foundation of Fujian Province, China, No. 2020R1011003 and No. 2022R1012001; and the Talents Training Project for the Key Young Scholars of Fujian Provincial Health Commission, China, No. 2021GGA056.
Institutional review board statement: The study was reviewed and approved by the Ethics Committee of Fujian Provincial Hospital (approval number: K2021 04 020).
Institutional animal care and use committee statement: All procedures involving animals were reviewed and approved by the Ethics Committee of Fujian Academy of Medical Sciences (approval number: DL2021-05).
Conflict-of-interest statement: All the authors report no relevant conflicts of interest for this article.
ARRIVE guidelines statement: The authors have read the ARRIVE guidelines, and the manuscript was prepared and revised according to the ARRIVE guidelines.
Data sharing statement: No additional data are available.
Open Access: This article is an open-access article that was selected by an in-house editor and fully peer-reviewed by external reviewers. It is distributed in accordance with the Creative Commons Attribution NonCommercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited and the use is non-commercial. See: https://creativecommons.org/Licenses/by-nc/4.0/
Corresponding author: Kun Wang, PhD, Associate Professor, Fujian Key Laboratory of Medical Analysis, Fujian Academy of Medical Sciences, No. 7 Wusi Road, Fuzhou 350001, Fujian Province, China. wangkun973@fjms.ac.cn
Received: December 28, 2024
Revised: March 26, 2025
Accepted: June 18, 2025
Published online: July 26, 2025
Processing time: 210 Days and 0.3 Hours
Abstract
BACKGROUND

Type 1 diabetes (T1D) results from the autoimmune-mediated loss of pancreatic β-cells. Current insulin therapies offer symptomatic relief but fall short of providing a definitive cure. Islet cell transplantation, while promising, faces limitations related to donor scarcity, procedural complexities, and the necessity for long-term immunosuppression. Consequently, there is an urgent need for innovative strategies aimed at β-cell regeneration. Patient-derived induced pluripotent stem cells (iPSCs), obtained from peripheral blood mononuclear cells (PBMCs) of T1D patients, hold great potential as a source of cells for therapeutic purposes. Therefore, the differentiation of T1D-iPSCs into functional pancreatic β-cells is a critical step toward effective β-cell replacement therapy.

AIM

To assess the potential of patient-derived T1D-β-like cells (differentiated from T1D-iPSCs reprogrammed from T1D-PBMCs) for restoring β-cell function in T1D.

METHODS

T1D-iPSCs were reprogrammed from T1D-PBMCs using an episomal vector-based approach. Pluripotency was confirmed by flow cytometry (FCM), quantitative real-time polymerase chain reaction, genomic stability analysis, and teratoma formation assays. Differentiation into pancreatic β-cells was optimized using triiodothyronine (T3), vitamin C (Vc), and an adenovirus (M3C) encoding pancreatic duodenal homeobox-1, neurogenin 3 (Ngn3), and MAF bZIP transcription factor A (MafA). Following characterization of β-cell features by immunofluorescence, quantitative real-time polymerase chain reaction, and flow cytometry, therapeutic efficacy was assessed through blood glucose monitoring after transplantation under the renal capsule of streptozotocin-induced diabetic mice.

RESULTS

T1D-iPSCs were successfully generated from T1D-PBMCs. These cells exhibited the hallmark characteristics of pluripotent stem cells, including appropriate morphology, differentiation potential, genomic integrity, and expression of pluripotency-associated genes. Differentiation yielded insulin-positive (insulin+) pancreatic β-like cells that, at the mRNA level, expressed key β-cell markers such as pancreatic duodenal homeobox-1, Ngn3, MafA, NeuroD, glucagon-like peptide-1 receptor, Nkx6.1, glucose transporter 2, and Kir6.2. Notably, the T3 + Vc group displayed the lowest Ngn3 expression (1.31 ± 0.38 vs 1.96 ± 0.25 vs 2.51 ± 0.24, P < 0.01), while the M3C + T3 + Vc group exhibited the highest MafA expression (0.49 ± 0.11 vs 0.32 ± 0.06 vs 0.29 ± 0.08, P < 0.05). Both in vitro and in vivo assessments confirmed the insulin secretion ability of the generated β-like cells; however, they did not demonstrate appropriate modulation of insulin release in response to variations in extracellular glucose concentrations.

CONCLUSION

T1D-iPSCs derived from T1D-PBMCs can be differentiated into insulin+ β-like cells, albeit with functional immaturity. These cells represent a potential source of seed cells for β-cell replacement therapy in T1D.

Keywords: Pluripotent stem cells; Cell reprogramming; Type 1 diabetes; Triiodothyronine; Vitamin C; Adenovirus; β-cell regeneration; Differentiation

Core Tip: This study successfully employed an episomal plasmid system to reprogram patient peripheral blood mononuclear cells into type 1 diabetes (T1D)-induced pluripotent stem cells (iPSCs). Optimization of the differentiation protocol, utilizing triiodothyronine (T3), vitamin C (Vc), and adenovirus-M3C, facilitated the directed differentiation of T1D-iPSCs toward pancreatic β-cells. The T3 + Vc group exhibited minimal neurogenin 3 expression, while the M3C + T3 + Vc group demonstrated maximal MAF bZIP transcription factor A expression. While insulin⁺ β-like cells differentiated from T1D-iPSCs displayed basal insulin secretion, they lacked the ability to respond to glucose stimulation, indicating functional immaturity compared to fully mature β-cells. These results underscore their potential as seed cells for β-cell replacement therapy in T1D.