Efficient derivation of microglia-like cells from human pluripotent stem cells

Abstract

Microglia, the only lifelong resident immune cells of the central nervous system (CNS), are highly specialized macrophages which have been recognized to play a crucial role in neurodegenerative diseases such as Alzheimer’s, Parkinson’s and Adrenoleukodystrophy (ALD). However, in contrast to other cell types of the human CNS, bona fide microglia have not yet been derived from cultured human pluripotent stem cells. Here we establish a robust and efficient protocol for the rapid production of microglia-like cells from human embryonic stem (ES) and induced pluripotent stem (iPS) cells that uses defined serum-free culture conditions. These in vitro pluripotent stem cell-derived microglia-like cells (termed pMGLs) faithfully recapitulate the expected ontogeny and characteristics of their in vivo counterparts and resemble primary fetal human and mouse microglia. We generated these cells from multiple disease-specific cell lines, and find that pMGLs derived from MeCP2 mutant hES cells are smaller than their isogenic controls. We further describe a culture platform to study integration and live behavior of pMGLs in organotypic 3D-cultures. This modular differentiation system allows the study of microglia in highly defined conditions, as they mature in response to developmentally relevant cues, and provides a framework to study the long-term interactions of microglia residing in a tissue-like environment.

Stem cell therapy for abrogating stroke-induced neuroinflammation and relevant secondary cell death mechanisms

“…Induced pluripotent stem cells are an attractive candidates for the treatment of neurodegenerative disorders due to their capacity to differentiate into a wide array of relevant phenotypes, including neural precursor cells, neurons, and vascular endothelial cells, upon addition of induction factors such as retinoic acid (Yuan et al, 2013; Muffat et al, 2016). iPSC neural progenitor cells have been found to spontaneously differentiate into neurons and astrocytes in vitro , expressing high levels of β-tubulin and glial fibrillary acidic protein (GFAP) (Paşca et al, 2015).…”

“…In terms of neuroinflammation, iPSCs may play a significant role. A recent in vitro study showed that iPSCs could differentiate into microglia, the primary immunoregulatory cell type in the brain (Muffat et al 2016). Moreover, a study by Hague and colleagues found that iPSCs could develop into antigen-specific regulatory T cells (Tregs), a T cell type involved in the suppression of autoimmunity (Haque et al, 2016).…”

Potassium channel expression and function in microglia: Plasticity and possible species variations

“…as in mouse microglia. Finally, we believe that the recent success in obtaining microglia-like cells from human pluripotent stem cells 51 should greatly aid this translational work, and even make it possible to investigate changes in channel expression in a broader disease specific context.…”

Pluripotent stem cell-derived organoids: using principles of developmental biology to grow human tissues in a dish

“…Microglia, which arise via primitive myelopoiesis, are also essential in neuronal maturation and development of the central nervous system. A recent report described the generation of microglia-like cells from human PSCs that integrate into a 3D organotypic neuroglial environment and respond to cellular damage (Muffat et al, 2016). Incorporation of PSC-derived microglia into cerebral organoids will provide an unprecedented opportunity to study the interactions between primitive myeloid-derived cells and early brain development.…”