Genetic programming of human iPSC-derived macrophages provides a tool to study the Erythroblastic Island niche in vitro
type: CreativeWork
description: Macrophages have been attracting much attention as they are present in many tissues and organs, and are involved in homeostatic tissue function and disease. Studies involving human macrophages have been hampered due to the ethical constraints and technical difficulties related to their isolation/derivation for in vitro expansion, and to the difficulty of genetically modifying them to interrogate the role of specific factors in macrophage behaviour. Thus, an off-the-shelf source of macrophages that is amenable to the vast arsenal of genetic manipulation techniques, such as induced pluripotent stem cells (iPSCs), represents a valuable tool in the macrophage field. As a proof of principle, we used a human iPSC-line that was targeted in the safe harbour AAVS1 locus to express the fluorescent protein ZsGreen constitutively (SFCi55-ZsG). We demonstrate efficient production of terminally differentiated macrophages from the SFCi55-ZsG iPSC-line that fluoresce green. Macrophages derived from this targeted cell line are indistinguishable from those generated from their parental line in terms of gene expression, cell surface marker expression and phagocytic ability. Furthermore, genetically modified macrophages could be activated/polarised to an M (LPS+ IFN-ϒ), M (IL10) or M (IL4) phenotype and retained their plasticity-related abilities, as they were able to switch from one activated state to another. These results showed that targeting of iPSCs via the AAVS1 locus allows for the production of fully functional genetically engineered macrophages that could be used for in vivo tracking of macrophages in disease. Furthermore, this strategy provides a platform for the introduction of genes/factors that are predicted to modulate and/or stabilise macrophage phenotype and function in diverse biological settings. We then used the AAVS1 targeting strategy to programme iPSC-derived macrophages into a phenotype comparable to macrophages associated with the erythroid island. Human red bl…
abstract: Macrophages have been attracting much attention as they are present in many tissues and organs, and are involved in homeostatic tissue function and disease. Studies involving human macrophages have been hampered due to the ethical constraints and technical difficulties related to their isolation/derivation for in vitro expansion, and to the difficulty of genetically modifying them to interrogate the role of specific factors in macrophage behaviour. Thus, an off-the-shelf source of macrophages that is amenable to the vast arsenal of genetic manipulation techniques, such as induced pluripotent stem cells (iPSCs), represents a valuable tool in the macrophage field. As a proof of principle, we used a human iPSC-line that was targeted in the safe harbour AAVS1 locus to express the fluorescent protein ZsGreen constitutively (SFCi55-ZsG). We demonstrate efficient production of terminally differentiated macrophages from the SFCi55-ZsG iPSC-line that fluoresce green. Macrophages derived from this targeted cell line are indistinguishable from those generated from their parental line in terms of gene expression, cell surface marker expression and phagocytic ability. Furthermore, genetically modified macrophages could be activated/polarised to an M (LPS+ IFN-ϒ), M (IL10) or M (IL4) phenotype and retained their plasticity-related abilities, as they were able to switch from one activated state to another. These results showed that targeting of iPSCs via the AAVS1 locus allows for the production of fully functional genetically engineered macrophages that could be used for in vivo tracking of macrophages in disease. Furthermore, this strategy provides a platform for the introduction of genes/factors that are predicted to modulate and/or stabilise macrophage phenotype and function in diverse biological settings. We then used the AAVS1 targeting strategy to programme iPSC-derived macrophages into a phenotype comparable to macrophages associated with the erythroid island. Human red bl…
author: Lopez Yrigoyen, Martha Beatriz
language: en
keywords: macrophages, erythroid island macrophage, KLF1 gene, red blood cell development
date published: 2019-11-25
date modified: 2026-07-24T02:14:03Z
provider: University of Edinburgh
publisher: Networked Digital Library of Theses and Dissertations
type: WebPage
source organization: The University of Edinburgh

PropertyValue
property ID: Handle
value: https://hdl.handle.net/1842/36580