Differentiation of Human induced Pluripotent Stem Cells to Authentic Macrophages using Defined, Serum Free, Open Source Media

Human iPSC and macrophages derived from them are increasingly popular tools for research into both infectious and degenerative diseases. However, as the field strives for greater modelling accuracy, it is becoming ever more challenging to justify the use of undefined and proprietary media for the culture of these cells. We describe here a defined, serum-free, open-source medium for the differentiation of iPSC-derived macrophages. This medium is equally capable of maintaining these cells compared to commercial alternatives. The macrophages differentiated in this medium display improved terminally differentiated cell characteristics, reduced basal expression of induced anti-viral response genes, and improved polarisation capacity. We conclude that cells cultured in this medium are an appropriate and malleable model for tissue resident macrophages, on which future differentiation techniques can be built. Overall design: We generated human iPSC-macrophages from the SFC840-03-03 cell line in two different media conditions. Macrophage precursor cells were harvested after 8, 10, and 12 weeks of differentiation and cultured for a further 7 days in terminal differentiation medium to make authentic macrophages. At this point cells were lysed for collection of RNA for RNA-Seq.