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However, our knowledge about the safe usage of anti-inflammatory drugs during pregnancy is still limited. While human induced pluripotent stem cells (hIPSC)-derived brain organoid models have started to explore functional consequences upon viral stimulation, these models commonly lack microglia, which are susceptible to and promote inflammation. Furthermore, microglia are actively involved in neuronal development. Here, we generate hIPSC-derived microglia precursor cells and assemble them into retinal organoids. Once the outer plexiform layer forms, these hIPSC-derived microglia (iMG) fully integrate into the retinal organoids. Since the ganglion cell survival declines by this time in 3D-retinal organoids, we adapted the model into 2D and identify that the improved ganglion cell number significantly decreases only with iMG presence. In parallel, we applied the immunostimulant POLY(I:C) to mimic a fetal viral infection. While POLY(I:C) exposure alters the iMG phenotype, it does not hinder their interaction with ganglion cells. Furthermore, iMG significantly enhance the supernatant\u2019s inflammatory secretome and increase retinal cell proliferation. Simultaneous exposure with the non-steroidal anti-inflammatory drug (NSAID) ibuprofen dampens POLY(I:C)-mediated changes of the iMG phenotype and ameliorates cell proliferation. Remarkably, while POLY(I:C) disrupts neuronal calcium dynamics independent of iMG, ibuprofen rescues this effect only if iMG are present. Mechanistically, ibuprofen targets the enzymes cyclooxygenase 1 and 2 (\n                    <jats:italic>COX1\/PTGS1<\/jats:italic>\n                    and\n                    <jats:italic>COX2\/PTGS2<\/jats:italic>\n                    ) simultaneously, from which iMG mainly express\n                    <jats:italic>COX1<\/jats:italic>\n                    . Selective\n                    <jats:italic>COX1<\/jats:italic>\n                    blockage fails to restore the calcium peak amplitude upon POLY(I:C) stimulation, suggesting ibuprofen\u2019s beneficial effect depends on the presence and interplay of\n                    <jats:italic>COX1<\/jats:italic>\n                    and\n                    <jats:italic>COX2<\/jats:italic>\n                    . These findings underscore the importance of microglia in the context of prenatal immune challenges and provide insight into the mechanisms by which ibuprofen exerts its protective effects during embryonic development.\n                  <\/jats:p>","DOI":"10.1186\/s12974-025-03366-x","type":"journal-article","created":{"date-parts":[[2025,4,4]],"date-time":"2025-04-04T04:41:48Z","timestamp":1743741708000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":5,"title":["Microglia determine an immune-challenged environment and facilitate ibuprofen action in human retinal organoids"],"prefix":"10.1186","volume":"22","author":[{"given":"Verena","family":"Schmied","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Medina","family":"Korkut-Demirba\u015f","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alessandro","family":"Venturino","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Juan Pablo","family":"Maya-Arteaga","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sandra","family":"Siegert","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2025,4,3]]},"reference":[{"key":"3366_CR1","doi-asserted-by":"publisher","first-page":"793","DOI":"10.1016\/J.TINS.2019.08.001","volume":"42","author":"U Meyer","year":"2019","unstructured":"Meyer U. Neurodevelopmental resilience and susceptibility to maternal immune activation. Trends Neurosci. 2019;42:793\u2013806. https:\/\/doi.org\/10.1016\/J.TINS.2019.08.001.","journal-title":"Trends Neurosci"},{"key":"3366_CR2","doi-asserted-by":"publisher","DOI":"10.1016\/J.BBADIS.2021.166198","volume":"1867","author":"C Auriti","year":"2021","unstructured":"Auriti C, De Rose DU, Santisi A, et al. Pregnancy and viral infections: mechanisms of fetal damage, diagnosis and prevention of neonatal adverse outcomes from cytomegalovirus to SARS-CoV-2 and Zika virus. Biochim Biophys Acta Mol Basis Dis. 2021;1867: 166198. https:\/\/doi.org\/10.1016\/J.BBADIS.2021.166198.","journal-title":"Biochim Biophys Acta Mol Basis Dis"},{"key":"3366_CR3","doi-asserted-by":"publisher","DOI":"10.3389\/FCIMB.2020.585727","volume":"10","author":"VS de Campos","year":"2020","unstructured":"de Campos VS, Calaza KC, Adesse D. Implications of TORCH diseases in retinal development\u2014special focus on congenital toxoplasmosis. Front Cell Infect Microbiol. 2020;10: 585727. https:\/\/doi.org\/10.3389\/FCIMB.2020.585727.","journal-title":"Front Cell Infect Microbiol"},{"key":"3366_CR4","doi-asserted-by":"publisher","DOI":"10.1136\/adc.42.222.110","author":"JA Dudgeon","year":"1967","unstructured":"Dudgeon JA. Maternal rubella and its effect on the foetus. Rev Article Arch Dis Childh. 1967. https:\/\/doi.org\/10.1136\/adc.42.222.110.","journal-title":"Rev Article Arch Dis Childh"},{"key":"3366_CR5","doi-asserted-by":"publisher","first-page":"1315","DOI":"10.1111\/RISA.12264","volume":"36","author":"KM Thompson","year":"2016","unstructured":"Thompson KM, Simons EA, Badizadegan K, et al. Characterization of the risks of adverse outcomes following rubella infection in pregnancy. 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