Direct Comparison of Four Hematopoietic Differentiation Methods from Human Induced Pluripotent Stem Cells

• Published in: Stem cell reports Vol. 15; no. 3; pp. 735 - 748
• Main Authors: Tursky, Melinda L., Loi, To Ha, Artuz, Crisbel M., Alateeq, Suad, Wolvetang, Ernst J., Tao, Helen, Ma, David D., Molloy, Timothy J.
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 08.09.2020; Elsevier
• Subjects: cell differentiation; disease modeling; Down syndrome; erythroid progenitor cells; hematopoiesis; hematopoietic progenitor cells; hematopoietic stem cells; induced pluripotent stem cells; pluripotent stem cells; β-thalassemia; hematopoiesis; cell differentiation; hematopoietic progenitor cells; erythroid progenitor cells; pluripotent stem cells; disease modeling; induced pluripotent stem cells; Down syndrome; hematopoietic stem cells; β-thalassemia
• ISSN: 2213-6711; 2213-6711
• DOI: 10.1016/j.stemcr.2020.07.009

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. [Display omitted] •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.