A systematic characterization of microglia-like cell occurrence during retinal organoid differentiation

Bartalska, Katarina; Hübschmann, Verena; Korkut-Demirbaş, Medina; Cubero, Ryan John; Venturino, Alessandro; Rössler, Karl; Czech, Thomas; Siegert, Sandra

doi:10.1016/j.isci.2022.104580

Cited by 9 publications

(11 citation statements)

References 109 publications

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“… Note: Ideally, the antibodies used for hiPSC-derived microglia-like cells are cross-validated for their specificity in human tissue because several papers only refer to the specificity in mouse tissue. In Bartalska et al., 1 we have validated the above set of markers in human brain tissue. …”

Section: Step-by-step Methods Detailsmentioning

confidence: 88%

“… 7 , 8 , 25 However, when we analyzed non-isolated microglia embedded in mesenchymal tissue, we could not observe TMEM119 expression in microglia-like cells. 1 Due to this ambiguity, we decided against including TMEM119 as a selective marker. Note: Ideally, the antibodies used for hiPSC-derived microglia-like cells are cross-validated for their specificity in human tissue because several papers only refer to the specificity in mouse tissue.…”

Section: Step-by-step Methods Detailsmentioning

confidence: 99%

Section: Step-by-step Methods Detailsmentioning

confidence: 99%

“…For generating hiPSC-derived microglia-like cells from hESCs or hiPSCs, we refer to. 1 , 4 , 5 , 6 , 7 , 8 , 9 , 10 , 11 , 12 , 13 , 14 , 15 , 16 The users can decide to either choose one strategy of the published protocols in Table 1 or buy a commercially available kit (e.g., STEMdiff™ Microglia Differentiation Kit). For a detailed step-by-step protocol, we refer to.…”

Section: Before You Beginmentioning

confidence: 99%

“…For this protocol, we used hiPSC-derived microglia-like cells that were collected from the supernatant from week 5 onwards twice per week as described in Bartalska et al. 1 At this time, the floating cells showed some small processes ( Figure 1 A). Note: This procedure depends on the chosen microglial differentiation protocol (see Table 1 ).…”

Section: Before You Beginmentioning

confidence: 99%

See 4 more Smart Citations

Assessing human iPSC-derived microglia identity and function by immunostaining, phagocytosis, calcium activity, and inflammation assay

Hübschmann

Korkut-Demirbaş²,

Siegert³

2022

STAR Protocols

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Section: Step-by-step Methods Detailsmentioning

confidence: 88%

Section: Step-by-step Methods Detailsmentioning

confidence: 99%

Section: Step-by-step Methods Detailsmentioning

confidence: 99%

Section: Before You Beginmentioning

confidence: 99%

Section: Before You Beginmentioning

confidence: 99%

See 3 more Smart Citations

Assessing human iPSC-derived microglia identity and function by immunostaining, phagocytosis, calcium activity, and inflammation assay

Hübschmann

Korkut-Demirbaş²,

Siegert³

2022

STAR Protocols

Self Cite

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Integrating human iPSC‐derived macrophage progenitors into retinal organoids to generate a mature retinal microglial niche

Usui‐Ouchi

Giles

Harkins‐Perry

et al. 2023

Glia

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In the retina, microglia are resident immune cells that are essential for development and function. Retinal microglia play a central role in mediating pathological degeneration in diseases such as glaucoma, retinitis pigmentosa, age‐related neurodegeneration, ischemic retinopathy, and diabetic retinopathy. Current models of mature human retinal organoids (ROs) derived from iPS cell (hiPSC) do not contain resident microglia integrated into retinal layers. Increasing cellular diversity in ROs by including resident microglia would more accurately represent the native retina and better model diseases in which microglia play a key role. In this study, we develop a new 3D in vitro tissue model of microglia‐containing retinal organoids by co‐culturing ROs and hiPSC‐derived macrophage precursor cells (MPCs). We optimized the parameters for successful integration of MPCs into retinal organoids. We show that while in the ROs, MPCs migrate to the equivalent of the outer plexiform layer where retinal microglia cells reside in healthy retinal tissue. While there, they develop a mature morphology characterized by small cell bodies and long branching processes which is only observed in vivo. During this maturation process these MPCs cycle through an activated phase followed by a stable mature microglial phase as seen by the down regulation of pro‐inflammatory cytokines and upregulation of anti‐inflammatory cytokines. Finally, we characterized mature ROs with integrated MPCs using RNAseq showing an enrichment of cell‐type specific microglia markers. We propose that this co‐culture system may be useful for understanding the pathogenesis of retinal diseases involving retinal microglia and for drug discovery directly in human tissue.

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Emerging Models to Study Human Microglia In vitro

Jäntti,

Kistemaker,

Buonfiglioli

et al. 2024

Advances in Neurobiology

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A systematic characterization of microglia-like cell occurrence during retinal organoid differentiation

Cited by 9 publications

References 109 publications

Assessing human iPSC-derived microglia identity and function by immunostaining, phagocytosis, calcium activity, and inflammation assay

Assessing human iPSC-derived microglia identity and function by immunostaining, phagocytosis, calcium activity, and inflammation assay

Integrating human iPSC‐derived macrophage progenitors into retinal organoids to generate a mature retinal microglial niche

Emerging Models to Study Human Microglia In vitro

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