High-Yield Human Induced Pluripotent Stem Cell-Derived Monocytes and Macrophages Are Functionally Comparable With Primary Cells

Abstract

Macrophages are pivotal effectors of host immunity and regulators of tissue homeostasis. Understanding of human macrophage biology has been hampered by the lack of reliable and scalable models for cellular and genetic studies. Human induced pluripotent stem cell (hiPSC)-derived monocytes and macrophages, as an unlimited source of subject genotype-specific cells, will undoubtedly play an important role in advancing our understanding of macrophage biology and implication in human diseases. In this study, we present a fully optimized differentiation protocol of hiPSC-derived monocytes and granulocyte-macrophage colony-stimulating factor (GM-CSF) or macrophage colony-stimulating factor (M-CSF). We present characterization of iPSC-derived myeloid lineage cells at phenotypic, functional, and transcriptomic levels, in comparison with corresponding subsets of peripheral blood-derived cells. We also highlight the application of hiPSC-derived monocytes and macrophages as a gene-editing platform for functional validation in research and drug screening, and the study also provides a reference for cell therapies.

Optimizing the Method for Differentiation of Macrophages from Human Induced Pluripotent Stem Cells

“…Recently, Gutbier et al modified the protocol of van Wilgenburg et al [ 21 ], and the calculated yield per iPSC (by one month) was comparable to that of our protocol (Table S1 ) [ 36 ]. Di Cui et al developed a high-yield monolayer differentiation protocol for about 2 × 10 4 monocytes per seeded hiPSC [ 30 ]. The significantly larger scale in cell production and efficient cryopreservation partially compensated for the other limitations of monolayer cultivation.…”

“…Compared with monolayer cultivation protocols, EB protocols have the advantage of allowing a continuous harvest of suspension cells for the long-term production of macrophages with relatively few cytokines [21][22][23][24][25][26]. By contrast, monolayer cultivation protocols achieved higher one-off collection rates of macrophage by modified differentiation steps, such as a complex mixture of factors [27][28][29][30] (up to 11 factors), hypoxia conditions [28], and CD14 sorting [29,30]. In the past, the seeding number of iPSCs per EB in most EB-based protocols was unfixed.…”

Quantitative Proteomics of Polarised Macrophages Derived from Induced Pluripotent Stem Cells

“…More recently, iPSC-derived M Φ (iPSDM) have emerged as a valuable platform for the generation of unlimited numbers of M Φ that are functionally similar to MDM [ 4 ]. In addition, the genetic manipulation of iPSCs to knock out genes of interest has paved the way for a better understanding of M Φ biology and has provided a platform for testing drug concepts that interfere with M Φ function [ 5 , 6 ].…”

Induced Pluripotent Stem Cell-Derived Monocytes/Macrophages in Autoinflammatory Diseases

“…To overcome the limitations described above, several methods have been developed to generate macrophages from PSCs. In this approach, the culture conditions drive embryonic stem cells or iPSCs to differentiate through a pathway that recapitulates embryonic hematopoiesis (23)(24)(25)(26). The advantages of PSC-MPs include easy availability, scalability, standardizability, and easy genetic manipulation (27,28).…”