Incorporation of stem cell-derived astrocytes into neuronal organoids to allow neuro-glial interactions in toxicological studies

Brüll, Markus; Spreng, Anna‐Sophie; Gutbier, Simon; Loser, Dominik; Krebs, A. T.; Reich, Marvin; Kraushaar, Udo; Britschgi, Markus; Patsch, Christoph; Leist, Marcel

doi:10.14573/altex.1911111

Cited by 26 publications

(35 citation statements)

References 107 publications

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“…This has fundamentally changed with the advent of stem cell-based protocols. We and others have already demonstrated the feasibility and applicability of iPSC-derived myeloid cells for the generation and study of microglia-like cells in 2D and 3D cultures [ 9 , 18 , 47 , 48 ]. Thus, these methods will improve macrophage research and drug discovery while reducing the need for animals for the isolation of primary cells.…”

Section: Discussionmentioning

confidence: 99%

Large-Scale Production of Human iPSC-Derived Macrophages for Drug Screening

Gutbier

Wanke

Dahm

et al. 2020

IJMS

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Tissue-resident macrophages are key players in inflammatory processes, and their activation and functionality are crucial in health and disease. Numerous diseases are associated with alterations in homeostasis or dysregulation of the innate immune system, including allergic reactions, autoimmune diseases, and cancer. Macrophages are a prime target for drug discovery due to their major regulatory role in health and disease. Currently, the main sources of macrophages used for therapeutic compound screening are primary cells isolated from blood or tissue or immortalized or neoplastic cell lines (e.g., THP-1). Here, we describe an improved method to employ induced pluripotent stem cells (iPSCs) for the high-yield, large-scale production of cells resembling tissue-resident macrophages. For this, iPSC-derived macrophage-like cells are thoroughly characterized to confirm their cell identity and thus their suitability for drug screening purposes. These iPSC-derived macrophages show strong cellular identity with primary macrophages and recapitulate key functional characteristics, including cytokine release, phagocytosis, and chemotaxis. Furthermore, we demonstrate that genetic modifications can be readily introduced at the macrophage-like progenitor stage in order to interrogate drug target-relevant pathways. In summary, this novel method overcomes previous shortcomings with primary and leukemic cells and facilitates large-scale production of genetically modified iPSC-derived macrophages for drug screening applications.

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Section: Discussionmentioning

confidence: 99%

Large-Scale Production of Human iPSC-Derived Macrophages for Drug Screening

Gutbier

Wanke

Dahm

et al. 2020

IJMS

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“…Simpler test systems were combined to obtain advanced complex test systems to better resemble the in vivo situation. These approaches include, for instance, the use of mixed cell cultures (Gutbier et al 2018 ), 3D organoids and neurospheres (Brull et al 2020 ; Hiemstra et al 2019 ; Kobolak et al 2020 ) and microphysiological systems (MPS), e.g. four-organ-chips developed to interconnect miniaturized human intestine, liver, brain and kidney equivalents (Ramme et al 2019 ) and improved iPSC cell differentiation to hepatocytes (Boon et al 2020 ).…”

Section: The Repository Of Nams Available For Nam-enhanced Raxmentioning

confidence: 99%

Setting the stage for next-generation risk assessment with non-animal approaches: the EU-ToxRisk project experience

et al. 2020

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In 2016, the European Commission launched the EU-ToxRisk research project to develop and promote animal-free approaches in toxicology. The 36 partners of this consortium used in vitro and in silico methods in the context of case studies (CSs). These CSs included both compounds with a highly defined target (e.g. mitochondrial respiratory chain inhibitors) as well as compounds with poorly defined molecular initiation events (e.g. short-chain branched carboxylic acids). The initial project focus was on developing a science-based strategy for read-across (RAx) as an animal-free approach in chemical risk assessment. Moreover, seamless incorporation of new approach method (NAM) data into this process (= NAM-enhanced RAx) was explored. Here, the EU-ToxRisk consortium has collated its scientific and regulatory learnings from this particular project objective. For all CSs, a mechanistic hypothesis (in the form of an adverse outcome pathway) guided the safety evaluation. ADME data were generated from NAMs and used for comprehensive physiological-based kinetic modelling. Quality assurance and data management were optimized in parallel. Scientific and Regulatory Advisory Boards played a vital role in assessing the practical applicability of the new approaches. In a next step, external stakeholders evaluated the usefulness of NAMs in the context of RAx CSs for regulatory acceptance. For instance, the CSs were included in the OECD CS portfolio for the Integrated Approach to Testing and Assessment project. Feedback from regulators and other stakeholders was collected at several stages. Future chemical safety science projects can draw from this experience to implement systems toxicology-guided, animal-free next-generation risk assessment.

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“…Although modifying the differentiation protocol results in innate microglia, there is also an increase in organoid variability with differences in microglial distribution between organoids [ 104 ]. The lack of microglia and reduced astrocytes in COs can be overcome by transplantation of human astrocytes and microglia derived from hiPSCs [ 105 – 108 ] into COs, which enhances the frequency of synchronized burst activity and CO development. This type of co-culture is stable long-term and provides a platform to model neuroinflammation and neuron-glia interactions.…”

Section: Challenges/limitationsmentioning

confidence: 99%