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悬浮培养改善iPSC扩张和多能性表型

Nerea Cuesta-Gomez^1,2, Kevin Verhoeff^1,2, Nidheesh Dadheech^3,4

¹Alberta Diabetes Institute, University of Alberta, Edmonton, AB, T6G 2T9, Canada.

Stem cell research & therapy

|June 6, 2023

概括

这项研究展示了一种新的3D Vertical-Wheel®生物反应器协议,用于扩展诱导多能干细胞 (iPSCs),在5天内实现近100倍的增长. 这种方法提高了细胞质量,并为再生药制造提供了成本高效的解决方案.

科学领域:

干细胞生物学干细胞生物学
再生医学是一种再生医学.
生物技术是生物技术.

背景情况:

诱导多能干细胞 (iPSC) 对再生医学至关重要,为各种细胞类型提供可再生来源.
高质量的人类iPSC的高效和成本效益的大规模制造对于临床翻译至关重要.
目前的方法在可扩展性和维持细胞质量方面面临挑战.

研究的目的:

为人类iPSC扩张提供一个改进的三维 (3D) Vertical-Wheel®生物反应器悬浮培养协议.
将3D悬浮协议与传统的二维 (2D) 平面培养方法进行比较.
为了评估在两种条件下培养的iPSCs的细胞扩张能力,遗传完整性和多能性.

主要方法:

人类外周血液单核细胞通过仙台病毒传染被重新编程成iPSC.
使用2D平面和3D垂直轮®生物反应器悬浮系统培养iPSC.
对比分析包括细胞扩张,基因完整性 (q-PCR),多能性标记物 (流细胞计),以及体外/体内分化潜力.

主要成果:

3D Vertical-Wheel®生物反应器在5天内实现了93.8倍的iPSC扩张,明显高于2D培养中的19.1倍 (p < 0.0022).
3D培养的iPSCs表现出增加的增殖 (Ki67+表达) 和更高频率的多能性标记物 (Oct4+Nanog+Sox2+).
基因分析证实了3D扩展的iPSC中没有常见突变,它也显示了从原始化过渡到原始化的多能性,并在体内产生了更成熟的瘤.

结论:

3D Vertical-Wheel®生物反应器协议实现了前所未有的iPSC扩张 (在5天内几乎是100倍),超过了之前的报道.
3D培养的iPSC显示出增强的体外和体外多能性,这表明质量有所改善.
这种可扩展和具有成本效益的3D培养系统对推进再生医学疗法和临床实施具有重大前景.

关键词:

生物反应器是一种生物反应器.细胞疗法是一种细胞疗法.扩张扩张扩张人类诱导的多能干细胞干细胞.多能性是一种多能性.干细胞是一种干细胞.这就是 iPSC 的意义.

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