Here, we present a protocol for generating long-term microglia-containing air-liquid-interface cortical organoid (MG-ALI-CO) cultures. This approach minimizes necrotic core formation, a common limitation of extended organoid cultures, favoring microglia survival and homeostasis. We describe steps for generating air-liquid-interface cortical organoids (ALI-COs), integrating macrophage precursors, and maintaining MG-ALI-COs. Additionally, we outline several experimental analyses of MG-ALI-COs, including immunostaining, imaging, and patch-clamp electrophysiological recordings. This model provides a physiologically relevant system to investigate human neuroimmune interactions in a 3D brain-like environment.
Protocol for generating and using human iPSC-derived microglia-containing air-liquid-interface cortical organoid cultures.
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作者：Cañizares Luna Marta, Mars Mayte, Jakobs Channa E, Huffels Christiaan F M, Ermakov Arthur, Hol Elly M, Pasterkamp R Jeroen
| 期刊： | STAR Protocols | 影响因子： | 1.300 |
| 时间： | 2025 | 起止号： | 2025 Jun 30; 6(3):103915 |
| doi： | 10.1016/j.xpro.2025.103915 | ||
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