Adapting human pluripotent stem cells to high-throughput and high-content screening

Area of Science:

• Stem cell biology
• Drug discovery and development
• High-throughput screening technologies

Background:

• Human pluripotent stem cells (hPSCs) are increasingly vital for drug discovery, cytotoxicity assessment, and disease modeling.
• Adapting hPSCs to large-scale culture and screening formats is crucial for their broader application.
• Existing methods may not be optimized for robust, high-throughput stem cell assays.

Purpose of the Study:

• To establish a simple and robust protocol for adapting human embryonic stem cells (hESCs) to high-throughput screening (HTS) formats.
• To enable the application of this protocol for human induced pluripotent stem cells (hiPSCs) and high-content screening (HCS).
• To develop a 7-day assay for identifying compounds affecting hESC self-renewal and differentiation.

Main Methods:

• Culture expansion of hESCs and adaptation to 384-well plates.
• Addition of small molecules or other factors for compound screening.
• Data acquisition and processing for assay analysis.
• Optimization of hESC plating density for 7-day HTS/HCS assays.

Main Results:

• A simple and robust protocol for adapting hESCs to HTS was successfully developed.
• The protocol is adaptable for hiPSCs and high-content screening (HCS).
• A 7-day assay was established to identify compounds influencing hESC self-renewal and differentiation.

Conclusions:

• The developed protocol facilitates the large-scale application of hPSCs in drug discovery and disease modeling.
• This adaptable assay provides a valuable tool for screening compounds affecting stem cell behavior.
• The optimized method supports efficient screening formats for stem cell-based research.