Correction to: Luminescent Human iPSC-Derived Neurospheroids Enable Modeling of Neurotoxicity After Oxygen–glucose Deprivation

Breedam, Elise Van; Nijak, Aleksandra; Buyle-Huybrecht, Tamariche; Stefano, Julia Di; Boeren, Marlies; Govaerts, Jonas; Quarta, Alessandra; Swartenbroekx, Tine; Jacobs, Eva; Menten, Björn; Gijsbers, Rik; Delputte, Peter; Alaerts, Maaike; Hassannia, Behrouz; Loeys, Bart; Berneman, Zwi; Timmermans, Jean‐Pierre; Berghe, Tom Vanden; Fransen, Erik; Wouters, An; Vos, Winnok H. De; Ponsaerts, Peter

doi:10.1007/s13311-022-01235-6

Cited by 2 publications

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“…hiPSC-neurons were generated from a previously established hiPSC-derived neural stem cell line (hiPSC-NSC) ( 25 ), whereby its differentiated progeny adopts a peripheral nervous system (PNS)-like neuronal phenotype after 3 weeks of culture ( Figure 1A ), as demonstrated by immunocytochemical analyses for NeuN, Tuj1 and Peripherin expression ( Figure 1B ). Obtained hiPSC-neuron cultures do not contain contaminating GFAP+ astrocytes ( Figure 1B ).…”

Section: Resultsmentioning

confidence: 99%

“…hiPSC-macrophages were generated from a previously established hiPSC line ( 25 ), the same that was used to generate the above described hiPSC-NSC line. Following hiPSC differentiation into hematopoietic progenitor cells (hiPSC-HPC) using a commercially available differentiation kit, a sequential expansion, and macrophage differentiation step ( Figure 3A ) was applied resulting in a population of round-shaped macrophage-like cells ( Figure 3B ).…”

Section: Resultsmentioning

confidence: 99%

“…A previously established hiPSC-derived neural stem cell line (hiPSC-NSC) ( 25 ) was differentiated to obtain Tuj1 + NeuN + peripherin + neurons, according to the protocol described in our earlier work ( 9 ). In brief, expanded hiPSC-NSC were grown onto poly-L-ornithine (20 µg/ml, Sigma) and laminin (10 µg/ml, Sigma)-coated well plates in neuronal differentiation medium (Neurobasal Plus medium (Gibco), 1 × B27 plus supplement (Gibco), 2 mM L-glutamine (Gibco), 1 × CultureOne supplement (Gibco), 200 µM ascorbic acid (Sigma), 1% PS (Gibco), 10 ng/ml recombinant human brain-derived neurotrophic factor (rhBDNF) and recombinant human glial cell line-derived neurotrophic factor (rhGDNF) (Immunotools)) for a minimum of 21 days.…”

Section: Methodsmentioning

confidence: 99%

“…hiPSC – the original hiPSC line from which hiPSC-NSC were derived ( 25 ) – were first differentiated into hematopoietic progenitor cells (hiPSC-HPC) using the STEMdiff Hematopoietic kit (STEMCELL Technologies) in 12 days, according to manufacturer’s instructions. At day 12, generated hiPSC-HPC were harvested and resuspended at a concentration of 3-5 x 10 5 cells/ml in StemSpan SFEMII medium (STEMCELL Technologies) containing 100 ng/ml SCF Immunotools), 100 ng/ml TPO (PeproTech), 100 ng/ml Flt-3l (Immunotools), 50 ng/ml M-CSF (Immunotools) and 20 ng/ml GM-CSF (Immunotools) to allow expansion of the cells.…”

Section: Methodsmentioning

confidence: 99%

“…Immunostaining of hiPSC-neurons was performed as previously described ( 25 ). In brief, cells were fixed with 4% paraformaldehyde for 20 min at 4°C and rinsed with PBS.…”

Section: Methodsmentioning

confidence: 99%

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Lack of strong innate immune reactivity renders macrophages alone unable to control productive Varicella-Zoster Virus infection in an isogenic human iPSC-derived neuronal co-culture model

Van Breedam,

Buyle-Huybrecht,

Govaerts

et al. 2023

Front. Immunol.

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With Varicella-Zoster Virus (VZV) being an exclusive human pathogen, human induced pluripotent stem cell (hiPSC)-derived neural cell culture models are an emerging tool to investigate VZV neuro-immune interactions. Using a compartmentalized hiPSC-derived neuronal model allowing axonal VZV infection, we previously demonstrated that paracrine interferon (IFN)-α2 signalling is required to activate a broad spectrum of interferon-stimulated genes able to counteract a productive VZV infection in hiPSC-neurons. In this new study, we now investigated whether innate immune signalling by VZV-challenged macrophages was able to orchestrate an antiviral immune response in VZV-infected hiPSC-neurons. In order to establish an isogenic hiPSC-neuron/hiPSC-macrophage co-culture model, hiPSC-macrophages were generated and characterised for phenotype, gene expression, cytokine production and phagocytic capacity. Even though immunological competence of hiPSC-macrophages was shown following stimulation with the poly(dA:dT) or treatment with IFN-α2, hiPSC-macrophages in co-culture with VZV-infected hiPSC-neurons were unable to mount an antiviral immune response capable of suppressing a productive neuronal VZV infection. Subsequently, a comprehensive RNA-Seq analysis confirmed the lack of strong immune responsiveness by hiPSC-neurons and hiPSC-macrophages upon, respectively, VZV infection or challenge. This may suggest the need of other cell types, like T-cells or other innate immune cells, to (co-)orchestrate an efficient antiviral immune response against VZV-infected neurons.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

“…Immunostaining of hiPSC-neurons was performed as previously described ( 25 ). In brief, cells were fixed with 4% paraformaldehyde for 20 min at 4°C and rinsed with PBS.…”

Section: Methodsmentioning

confidence: 99%

See 3 more Smart Citations

Lack of strong innate immune reactivity renders macrophages alone unable to control productive Varicella-Zoster Virus infection in an isogenic human iPSC-derived neuronal co-culture model

Van Breedam,

Buyle-Huybrecht,

Govaerts

et al. 2023

Front. Immunol.

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Varicella-zoster virus recapitulates its immune evasive behaviour in matured hiPSC-derived neurospheroids

Govaerts,

Van Breedam,

De Beuckeleer

et al. 2024

Front. Immunol.

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Varicella-zoster virus (VZV) encephalitis and meningitis are potential central nervous system (CNS) complications following primary VZV infection or reactivation. With Type-I interferon (IFN) signalling being an important first line cellular defence mechanism against VZV infection by the peripheral tissues, we here investigated the triggering of innate immune responses in a human neural-like environment. For this, we established and characterised 5-month matured hiPSC-derived neurospheroids (NSPHs) containing neurons and astrocytes. Subsequently, NSPHs were infected with reporter strains of VZV (VZVeGFP-ORF23) or Sendai virus (SeVeGFP), with the latter serving as an immune-activating positive control. Live cell and immunocytochemical analyses demonstrated VZVeGFP-ORF23 infection throughout the NSPHs, while SeVeGFP infection was limited to the outer NSPH border. Next, NanoString digital transcriptomics revealed that SeVeGFP-infected NSPHs activated a clear Type-I IFN response, while this was not the case in VZVeGFP-ORF23-infected NSPHs. Moreover, the latter displayed a strong suppression of genes related to IFN signalling and antigen presentation, as further demonstrated by suppression of IL-6 and CXCL10 production, failure to upregulate Type-I IFN activated anti-viral proteins (Mx1, IFIT2 and ISG15), as well as reduced expression of CD74, a key-protein in the MHC class II antigen presentation pathway. Finally, even though VZVeGFP-ORF23-infection seems to be immunologically ignored in NSPHs, its presence does result in the formation of stress granules upon long-term infection, as well as disruption of cellular integrity within the infected NSPHs. Concluding, in this study we demonstrate that 5-month matured hiPSC-derived NSPHs display functional innate immune reactivity towards SeV infection, and have the capacity to recapitulate the strong immune evasive behaviour towards VZV.

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Correction to: Luminescent Human iPSC-Derived Neurospheroids Enable Modeling of Neurotoxicity After Oxygen–glucose Deprivation

Abstract: The original article has been corrected.Publisher's Note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Cited by 2 publications

References 0 publications

Lack of strong innate immune reactivity renders macrophages alone unable to control productive Varicella-Zoster Virus infection in an isogenic human iPSC-derived neuronal co-culture model

Lack of strong innate immune reactivity renders macrophages alone unable to control productive Varicella-Zoster Virus infection in an isogenic human iPSC-derived neuronal co-culture model

Varicella-zoster virus recapitulates its immune evasive behaviour in matured hiPSC-derived neurospheroids

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