Abstract
Induced pluripotent stem cells (iPSCs) are an invaluable resource for the study of human disease. However, there are no standardized methods for differentiation into hematopoietic cells, and there is a lack of robust, direct comparisons of different methodologies. In the current study we improved a feeder-free, serum-free method for generation of hematopoietic cells from iPSCs, and directly compared this with three other commonly used strategies with respect to efficiency, repeatability, hands-on time, and cost. We also investigated their capability and sensitivity to model genetic hematopoietic disorders in cells derived from Down syndrome and β-thalassemia patients. Of these methods, a multistep monolayer-based method incorporating aryl hydrocarbon receptor hyperactivation (“2D-multistep”) was the most efficient, generating significantly higher numbers of CD34
+
progenitor cells and functional hematopoietic progenitors, while being the most time- and cost-effective and most accurately recapitulating phenotypes of Down syndrome and β-thalassemia.
•
Direct comparison of 4 serum & feeder-free iPSC hematopoietic differentiation methods
•
Comparison: cost-benefit efficiency, sensitivity to model genetic blood diseases
•
Presents an improved iPSC hematopoietic differentiation: 7× efficiency at 50% cost
•
Improved method = most live cells, CD34
+
, CFU; lowest cost; greatest sensitivity
Tursky and colleagues addressed a vital, unmet need to aid method selection for iPSC hematopoietic differentiation for research and potentially clinical use, comparing four representative serum- and feeder-layer-free methods. Assessments include production efficiency of CD34
+
and functional hematopoietic progenitor cells, cost-benefit analysis, and ability to recapitulate aberrant hematopoiesis. An optimized method proved the most efficient, cost-effective, and sensitive.