Scale up manufacturing approach for production of human induced pluripotent stem cell-derived islets using Vertical Wheel® bioreactors.

IF 6.4 1区医学 Q1 CELL & TISSUE ENGINEERING

npj Regenerative Medicine Pub Date : 2025-05-29 DOI:10.1038/s41536-025-00409-y

Nidheesh Dadheech, Mario Bermúdez de León, Zofia Czarnecka, Nerea Cuesta-Gomez, Ila Tewari Jasra, Rena Pawlick, Braulio Marfil-Garza, Sandhya Sapkota, Kevin Verhoeff, Haide Razavy, Perveen Anwar, Abhineet Singh, Nilanjan Ray, Doug O' Gorman, Glen Jickling, James Lyon, Patrick MacDonald, A M James Shapiro

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引用次数: 0

Abstract

Advanced protocols show potential for human stem cells (SC)-derived islets generation under planar (2D) alone or three-dimensional (3D) cultures, but show challenges in scalability, cell loss, and batch-to-batch consistency. This study explores Vertical Wheel (VW)® bioreactor suspension technology to differentiate islets from human induced pluripotent stem cells, achieving uniform, transcriptionally mature, and functional SC-islets. A 5x increase in scale from 0.1 L to 0.5 L reactors resulted in a 12-fold (15,005-183,002) increase in islet equivalent count (IEQ) without compromising islet structure. SC-islets show enriched β-cell composition (~63% CPPT⁺NKX6.1⁺ISL1⁺), glucose responsive insulin release (3.9-6.1-fold increase), and reversed diabetes in STZ-treated mice. Single cell RNA sequencing and flowcytometry analysis confirmed transcriptional maturity and functional identity, similar to adult islets. Lastly, harvested SC-islet grafts demonstrate improved islet functionality and mature transcriptomic signatures. Overall, scale-up in VW® bioreactor technology enhances IEQ yield with minimal variability and reduced cell loss, offering a pathway for clinical-grade SC-islet production.

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利用Vertical Wheel®生物反应器，扩大人类诱导多能干细胞衍生胰岛的生产规模。

先进的方案显示了在平面（2D）单独或三维（3D）培养下生成人类干细胞（SC）衍生胰岛的潜力，但在可扩展性、细胞损失和批间一致性方面存在挑战。本研究探索了垂直轮（VW）®生物反应器悬浮技术，将胰岛与人类诱导多能干细胞区分开来，获得统一的、转录成熟的、功能性的sc -胰岛。从0.1 L到0.5 L反应器的规模增加5倍，导致胰岛当量计数（IEQ）增加12倍（15,005-183,002），而不影响胰岛结构。sc -胰岛显示出丰富的β-细胞组成（~63% CPPT+NKX6.1+ISL1+），葡萄糖反应性胰岛素释放（增加3.9-6.1倍），并在stz治疗小鼠中逆转糖尿病。单细胞RNA测序和流式细胞术分析证实了转录成熟度和功能特性，与成人胰岛相似。最后，收获的sc -胰岛移植物显示出改善的胰岛功能和成熟的转录组特征。总体而言，VW®生物反应器技术的扩大提高了IEQ产量，减少了细胞损失，为临床级sc -胰岛的生产提供了一条途径。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

npj Regenerative Medicine Engineering-Biomedical Engineering

CiteScore

10.00

自引率

1.40%

发文量

审稿时长

12 weeks

期刊介绍： Regenerative Medicine, an innovative online-only journal, aims to advance research in the field of repairing and regenerating damaged tissues and organs within the human body. As a part of the prestigious Nature Partner Journals series and in partnership with ARMI, this high-quality, open access journal serves as a platform for scientists to explore effective therapies that harness the body's natural regenerative capabilities. With a focus on understanding the fundamental mechanisms of tissue damage and regeneration, npj Regenerative Medicine actively encourages studies that bridge the gap between basic research and clinical tissue repair strategies.