Integrated, automated maintenance, expansion and differentiation of 2D and 3D patient-derived cellular models for high throughput drug screening

Abstract

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.

Gene-corrected p.A30P SNCA patient-derived isogenic neurons rescue neuronal branching and function

“…Each directed neuronal differentiation took place using NPCs with different passage numbers (10 > p < 30), where the neurons were expanded and directly differentiated for a minimum period of 30 days. The generation and characterisation of the NPCs and vmDA neurons used this study has been described with the protocol elsewhere repeated and made automation compatible for high-throughput drug screening 30 , 67 , 72 .…”

Patient-Derived Organoids as a Model for Cancer Drug Discovery

“…Therefore, it appears to be feasible to employ organoids in these assays, and it is hoped that the modification of the culture conditions will allow the automation of the process, thus reducing the variability while further maximizing the assays performed on each sample. An automated platform for the high-throughput screening of patient-derived 3D organoids has recently been described; however, this system was not able to use organoids embedded in the extracellular matrix [82]. Using an automated platform, organoids derived from single cells from colon cancer tissue were established in a 384-well plate format which could be used for testing drugs in a high-throughput assay [30].…”

Genome Editing in iPSC-Based Neural Systems: From Disease Models to Future Therapeutic Strategies

“…Thus, arrayed screens usually entail the selection of a focused set of genes for perturbation when complex phenotypes are analyzed (Heman-Ackah et al, 2016 ; Deneault et al, 2018 ; Kwart et al, 2019 ), restraining the identification of disease mechanisms and/or new therapeutic targets to already known disease-associated variants. New advances in automated platforms for currently labor intensive iPSCs culture and complex differentiation protocols, both in 2D and 3D (Daily et al, 2017 ; Dhingra et al, 2020 ; Renner et al, 2020 ; Woodruff et al, 2020 ; Boussaad et al, 2021 ), will help in ensuring reproducibility of cellular phenotypes and high-throughput readouts. Technological development will in the future improve our ability to perform such screens and increase the number of variants that can be screened still further, expanding our knowledge about brain physiology and mechanisms that drive brain disorders with implications for therapeutic interventions.…”