Integrated, automated maintenance, expansion and differentiation of 2D and 3D patient-derived cellular models for high throughput drug screening.
Integrated, automated maintenance, expansion and differentiation of 2D and 3D patient-derived cellular models for high throughput drug screening.
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用于高通量药物筛选的2D和3D患者源性细胞模型的集成、自动化维护、扩展和分化。
DOI：
10.1038/s41598-021-81129-3
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发表时间：
2021-01-14
影响因子：
4.6
通讯作者：
Krüger R
中科院分区：
文献类型：
--
作者：
Boussaad I;Cruciani G;Bolognin S;Antony P;Dording CM;Kwon YJ;Heutink P;Fava E;Schwamborn JC;Krüger R
患者来源的细胞模型成为一种越来越强大的工具，可以为精准医学方法建模人类疾病。在这些模型中鉴定出稳健的细胞疾病表型为高通量筛选（HTS）铺平了道路，包括实验室高级自动化的实现。然而，用于HTS的细胞的维护和扩增仍然主要是手工工作。在这里，我们描述了一个综合的，复杂的自动化平台，HTS在转化研究设置也设计用于维护和扩展不同的细胞类型。全面的设计允许所有培养步骤的自动化，并灵活地开发用于不同细胞类型的方法。我们展示了控制细胞接种，分裂和人类成纤维细胞，诱导多能干细胞（iPSC）和神经祖细胞（NPC）的扩增协议，允许随后分化成不同的细胞类型和基于图像的多参数筛选。此外，我们提供了在2D培养中NPC神经元分化的自动化方案和用于HTS的3D中脑类器官。该多任务平台的灵活性使其成为转化研究环境的理想解决方案，涉及在不同患者来源的细胞模型上进行精准医学实验。
Patient-derived cellular models become an increasingly powerful tool to model human diseases for precision medicine approaches. The identification of robust cellular disease phenotypes in these models paved the way towards high throughput screenings (HTS) including the implementation of laboratory advanced automation. However, maintenance and expansion of cells for HTS remains largely manual work. Here, we describe an integrated, complex automated platform for HTS in a translational research setting also designed for maintenance and expansion of different cell types. The comprehensive design allows automation of all cultivation steps and is flexible for development of methods for variable cell types. We demonstrate protocols for controlled cell seeding, splitting and expansion of human fibroblasts, induced pluripotent stem cells (iPSC), and neural progenitor cells (NPC) that allow for subsequent differentiation into different cell types and image-based multiparametric screening. Furthermore, we provide automated protocols for neuronal differentiation of NPC in 2D culture and 3D midbrain organoids for HTS. The flexibility of this multitask platform makes it an ideal solution for translational research settings involving experiments on different patient-derived cellular models for precision medicine.
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影响因子：
4.7
作者：
Gribkoff VK;Kaczmarek LK
通讯作者：
Kaczmarek LK
影响因子：
5.2
作者：
Joannides, Alexis;Fiore-Heriche, Christelle;Chandran, Siddharthan
通讯作者：
Chandran, Siddharthan
影响因子：
1.2
作者：
Mitchell, Leslie A.;Phillips, Nick A.;Boeke, Jef D.
通讯作者：
Boeke, Jef D.
影响因子：
5.9
作者：
Phan, Nhan;Hong, Jenny J.;Soragni, Alice
通讯作者：
Soragni, Alice
诱导多能干细胞作为加速患者和疾病特异性药物发现的模型。
影响因子：
4.1
作者：
Gunaseeli I;Doss MX;Antzelevitch C;Hescheler J;Sachinidis A
通讯作者：
Sachinidis A