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优化和可扩展的人类外周血液单核细胞的无预涂层重编程,将其转化为iPSC

Elisabetta Fiacco¹, Sara Landi¹, Jacopo Zasso¹

¹Human Technopole, Milan, Italy.

Current protocols

|January 24, 2024

概括

一种新方法通过消除表面预涂层步骤来简化人类诱导的多能干细胞 (iPSC) 生成. 这项创新简化了用于疾病建模的成本效益高,可扩展和自动化的iPSC生产协议.

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.
细胞重新编程的细胞重编程.

背景情况:

人类诱导的多能干细胞 (iPSC) 对于疾病建模和生理学研究至关重要.
目前的iPSC生成协议通常是复杂的,耗时的,并且难以自动化.
培养表面的预涂层是iPSC培养中的标准但费力的步骤.

研究的目的:

开发一种简化,成本效益和可扩展的方法来生成iPSC.
为了消除 iPSC 重编程协议中对培养表面进行预涂层的需要.
为了促进自动化和高吞吐量iPSC生产.

主要方法:

开发了一种无预涂层的方法,用于重新编程血源单核细胞 (PBMC).
使用含有拉米宁-511碎片的培养基进行细胞播种.
优化细胞密度,培养基成分和iPSC传播技术.
整合了一个优化的子克隆方法,以提高可扩展性.

主要成果:

成功地重新编程新鲜或冷的PBMC,而不需要预涂表面.
该协议是自动化的友好,使高吞吐量处理.
减少与手工处理和涂装前步骤相关的劳动力和成本.
该方法适用于小型和大型iPSC生成.

结论:

开发的没有预涂层的方法大大简化了iPSC生成.
这种方法提高了iPSC生产的可扩展性,可复制性和成本效益.
该协议非常适合用于疾病建模和再生医学中的自动化管道和高通量应用.

关键词:

涂层涂层是一种涂层.细胞外矩阵是细胞外矩阵.诱导多能干细胞的诱导干细胞.拉米宁-51111可以使用.在重新编程PBMC时.缩小了规模,降低了规模.

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