Kayama Akihiro, Eto Koji
Department of Clinical Application, Center for iPS Cell Research and Application (CiRA), Kyoto University, 53 Shōgoin Kawarachō, Sakyo Ward, Kyoto, 606-8397, Japan.
Regen Ther. 2024 Jan 4;25:213-219. doi: 10.1016/j.reth.2023.12.009. eCollection 2024 Mar.
The production of platelets from induced pluripotent cells (iPSCs) may offer a safer and sustainable alternative for transfusions and drug delivery systems (DDS). However, the mass production of the clinically required number of iPSC-derived platelets (iPSC-PLTs) is challenging. Here, we introduce recent technologies for mass production and the first-in-human clinical trial using iPSC-PLTs. To this end, we established immortalized megakaryocyte progenitor cell lines (imMKCLs) as an expandable master cell bank (MCB) through the overexpression of c-MYC, BMI1 and BCL-XL, which modulated megakaryopoiesis and thrombopoiesis. We also optimized a culture cocktail for maturation of the imMKCLs by mixing an aryl hydrocarbon receptor (AhR) antagonist, SR1/GNF-316; a Rho-associated protein kinase (ROCK) inhibitor, Y-27632/Y-39983; and a small-molecule compound replacing recombinant thrombopoietin (TPO), TA-316. Finally, we discovered the importance of turbulence on the manufacturing of intact iPSC-PLTs, allowing us to develop a turbulence-based bioreactor, VerMES. Combination of the MCB and VerMES enabled us to produce more than 100 billion iPSC-PLTs, leading to the first-in-human clinical trial. Despite these advancements, many challenges remain before expanding the clinical implementation of this strategy.
从诱导多能干细胞(iPSC)生产血小板可能为输血和药物递送系统(DDS)提供一种更安全、可持续的替代方案。然而,大规模生产临床所需数量的iPSC来源的血小板(iPSC-PLT)具有挑战性。在此,我们介绍了用于大规模生产的最新技术以及使用iPSC-PLT的首次人体临床试验。为此,我们通过过表达c-MYC、BMI1和BCL-XL建立了永生化巨核细胞祖细胞系(imMKCL)作为可扩增的主细胞库(MCB),这些基因调节了巨核细胞生成和血小板生成。我们还通过混合芳烃受体(AhR)拮抗剂SR1/GNF-316、Rho相关蛋白激酶(ROCK)抑制剂Y-27632/Y-39983和替代重组血小板生成素(TPO)的小分子化合物TA-316,优化了用于imMKCL成熟的培养混合物。最后,我们发现了湍流在完整iPSC-PLT制造中的重要性,从而使我们能够开发基于湍流的生物反应器VerMES。MCB和VerMES的结合使我们能够生产超过1000亿个iPSC-PLT,从而促成了首次人体临床试验。尽管取得了这些进展,但在扩大该策略的临床应用之前,仍有许多挑战。