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

Di Cui,Sophie A Fillon,Bernd Weigle,Heinrich J Huber,Linda Jannetti,Katrin Fundel-Clemens,Timo Isambert,Alexander Ghanem,Alexandra Franz,Akira Niwa,Coralie Viollet,Stefan Pflanz

doi:10.3389/fcell.2021.656867

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.

Highlights

Monocytes and macrophages, as a central part of the host immune system, play essential roles in homeostasis and inflammation
Hematopoietic progenitor cells (HPCs) started to emerge as floaters around the organoid (Figure 1B, Day 6) in Stem line II medium supplemented with VEGF, stem cell factor (SCF), thyroid peroxidase (Tpo), interleukin (IL)-3, and FLT-3 Ligand (FL) from Day 4 to Day 6
HPCs were further differentiated into myeloid progenitors (MPs) (Figure 1B, Day 10) with additional macrophage colony-stimulating factor (M-CSF) as well as SCF, Tpo, IL-3, and FL in Stem line II medium for 4 days

Summary

INTRODUCTION

As a central part of the host immune system, play essential roles in homeostasis and inflammation. The commonly used resource for in vitro experiments with human cells is macrophages derived from monocytes isolated from donor blood, which has the advantage of using patientspecific cells regarding the pathophysiology of different diseases. The inter-donor variation is an important factor that can affect the reproducibility of results Another alternative way preferred by many researches is using the immortalized cells like THP-1 or U937 cell line (Rogers and Schwartz, 2018). Accumulating studies have shown the potential of hiPSC-derived monocytic like cells in disease-associated research and therapeutic modeling (Karlsson et al, 2008; Senju et al, 2011; Takamatsu et al, 2014; Haenseler et al, 2017; Ackermann et al, 2018; Hong et al, 2018). Our new protocol provides a reliable, scalable, and gene-editable system for human monocyte and macrophage research

RESULTS

DISCUSSION

EXPERIMENTAL PROCEDURES

ETHICS STATEMENT