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. 2023 Nov 22;13:20477. doi: 10.1038/s41598-023-46852-z

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© The Author(s) 2023

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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Q60 iPSC-microglia showed impairment in endocytosis and increased cytokine secretion compared to Q33 iPSC-microglia. (a) Confirmation of cell identity of both Q33 and Q60 iPSC-microglia using IBA1 and TMEM119. Scale bar = 50 μm. (b) Investigation of iPSC-microglia morphology showed no differences between Q33 and Q60 iPSC-microglia in cell area and roundness. (c) Unstimulated iPSC-microglia supernatants were assessed by ELISA for the secretion of the cytokines IL-8 and IL-6. Unstimulated Q60 iPSC-microglia showed an increased secretion of IL-6, but not IL-8. (d) Phagocytosis was assessed via the uptake of pHrodo-conjugated E. coli particles. Q33 and Q60 iPSC-microglia were imaged every 15 min for 4 h. Uptake of E. coli particles was inhibited by the addition of 30 μM cytochalasin D (CytoD) for 1 h prior and during phagocytosis. Total spot area of phagocytosed E. coli after 4 h was not different between Q33 and Q60 iPSC-microglia. (e) Endocytosis was assessed via the uptake of pHrodo-conjugated transferrin particles. Q33 and Q60 iPSC-microglia were imaged every 10 min for 4 h. Uptake of transferrin particles was inhibited by the addition of 30 μM dynasore for 1 h prior and during endocytosis. Total spot area of transferrin particles was significantly reduced in Q60 iPSC-microglia compared to Q33 control iPSC-microglia. Data are expressed as mean ± SEM. n = 3 independent experimental repeats with 1 clone. Statistical analysis was performed using unpaired t-test. *p < 0.05; ***p < 0.001.