使用Vertical Wheel®生物反应器扩大生产人诱导多能干细胞衍生胰岛的制造方法。

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

作者信息

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

机构信息

Alberta Diabetes Institute, University of Alberta, Edmonton, AB, Canada.

Department of Surgery, University of Alberta, Edmonton, AB, Canada.

出版信息

NPJ Regen Med. 2025 May 29;10(1):24. doi: 10.1038/s41536-025-00409-y.

DOI:10.1038/s41536-025-00409-y

PMID:40442082

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC12122946/

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% CPPTNKX6.1ISL1), 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.

摘要

先进的方案显示出在二维（2D）单独培养或三维（3D）培养下从人干细胞（SC）生成胰岛的潜力，但在可扩展性、细胞损失和批次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，以从人诱导多能干细胞分化生成胰岛，从而获得均匀、转录成熟且功能正常的SC胰岛。反应器规模从0.1升扩大到0.5升，扩大了5倍，胰岛当量计数（IEQ）增加了12倍（从15,005个增加到183,002个），同时不影响胰岛结构。SC胰岛显示出丰富的β细胞组成（约63% CPPTNKX6.1ISL1）、葡萄糖反应性胰岛素释放（增加3.9至6.1倍），并使链脲佐菌素处理的小鼠的糖尿病得到逆转。单细胞RNA测序和流式细胞术分析证实了其转录成熟度和功能特性，与成年胰岛相似。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®生物反应器技术的放大提高了IEQ产量，变异性最小且细胞损失减少，为临床级SC胰岛的生产提供了一条途径。