Protocol for generating megakaryocytes from patient induced pluripotent stem cells for disease modeling and compound screening

Here, we present a protocol for generating megakaryocytes from patient induced pluripotent stem cells (iPSCs). We describe steps for expanding and dissociating iPSCs into single cells and aggregating them into spin embryoid bodies (EBs). We detail procedures for lineage commitment and differentiation toward megakaryocytes within 14 days. This protocol supports phenotypic characterization and compound testing across multiple patient-derived iPSC lines. For complete details on the use and execution of this protocol, please refer to Flosdorf et al. 1 • Instructions for differentiating patient iPSCs into megakaryocytes • Steps for efficient mesoderm induction and cytokine- and compound-guided differentiation • Procedures for disease modeling and compound screening Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. Here, we present a protocol for generating megakaryocytes from patient induced pluripotent stem cells (iPSCs). We describe steps for expanding and dissociating iPSCs into single cells and aggregating them into spin embryoid bodies (EBs). We detail procedures for lineage commitment and differentiation toward megakaryocytes within 14 days. This protocol supports phenotypic characterization and compound testing across multiple patient-derived iPSC lines.