Home›Pharmacology›A Simple Method to Identify Kinases That Regulate Embryonic Stem Cell Pluripotency by High-throughput Inhibitor Screening
PharmacologyJoVE (Open Access)Citable · DOI
A Simple Method to Identify Kinases That Regulate Embryonic Stem Cell Pluripotency by High-throughput Inhibitor Screening
DOI: 10.3791/55515-v
What you'll learn
✓Design and execute a high-throughput kinase inhibitor screen on embryonic stem cells
✓Analyze screening data to identify kinase regulators of pluripotency transitions
✓Apply quantitative protocols to evaluate small molecule effects on stem cell fate
Protocol
Here, we present a quantitative and scalable protocol to perform targeted small molecule screens for kinase regulators of the naïve-primed pluripotent transition.
Difficulty
advanced
Total time
~5–7 days (including cell culture preparation, screening, and analysis)
Model organism
Mouse embryonic stem cells (mESC)
Biosafety
BSL-1
Steps
1
Prepare mouse embryonic stem cells for screening
Culture mESCs under defined conditions and prepare cell populations for kinase inhibitor screening, establishing baseline pluripotency state.
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2
Conduct kinase inhibitor screening and data analysis
Perform targeted small molecule screening against kinase library, apply quantitative readouts to assess pluripotency modulation across inhibitor conditions.