An Automated High Throughput Screening-Compatible Assay to Identify Regulators of Stem Cell Neural Differentiation

Abstract

The use of Embryonic Stem Cells (ESCs) holds considerable promise both for drug discovery programs and the treatment of degenerative disorders in regenerative medicine approaches. Nevertheless, the successful use of ESCs is still limited by the lack of efficient control of ESC self-renewal and differentiation capabilities. In this context, the possibility to modulate ESC biological properties and to obtain homogenous populations of correctly specified cells will help developing physiologically relevant screens, designed for the identification of stem cell modulators. Here, we developed a high throughput screening-suitable ESC neural differentiation assay by exploiting the Cell(maker) robotic platform and demonstrated that neural progenies can be generated from ESCs in complete automation, with high standards of accuracy and reliability. Moreover, we performed a pilot screening providing proof of concept that this assay allows the identification of regulators of ESC neural differentiation in full automation.

Emerging Requirements for Technology Management: A Sector-based Scenario Planning Approach

“…The development of new drug compounds continues to be a scientifi cally and technologically challenging area. Whilst the biotechnology business model has yielded some major innovations in the way drug compounds are developed and drug discovery processes, such as high-throughput screening (Casalino, 2012), have also helped in this regard. Nevertheless, pharmaceutical companies continue to make major levels of investment in scientifi c research and increasingly this is with external research suppliers.…”

HTS/HCS to Screen Molecules Able to Maintain Embryonic Stem Cell Self-Renewal or to Induce Differentiation: Overview of Protocols

“…The data presented let us to suggest that the method described by Casalino et al is easy to be reproduced, although it has until now never been used to screen a library of compounds [60]. Moreover, both methods proposed by Cainarca et al [60] and McNeisch et al [61] seem to be very efficient in the discovery of active molecules, even though the protocol described by McNeisch et al requires many manipulations and different growth factors to produce the neuronal precursors necessary for the screening.…”

“…Starting from a HTS compatible single-step protocol to differentiate ES cells into neuron and glial cells [12], a new HTS assay suitable for the identification of modulators of ES cell neural differentiation was developed, demonstrating that neural progenies can be generated from ES cells in complete automation, with high standards of accuracy and reliability [60]. A number of screenings have been performed on neuronal precursors pre-differentiated from ES cells.…”

“…Moreover, both methods proposed by Cainarca et al [60] and McNeisch et al [61] seem to be very efficient in the discovery of active molecules, even though the protocol described by McNeisch et al requires many manipulations and different growth factors to produce the neuronal precursors necessary for the screening.…”

Induced Pluripotent Stem Cells as a Disease Modeling and Drug Screening Platform

“…Further hurdles may depend on the particular features of iPSC growth and culture conditions, which can complicate their implementation in industrial scale screens [75]. Nevertheless, high-throughput screens in human pluripotent cells performed thus far demonstrate the effectiveness of these pioneer systems [122, 124–127]. …”