TY - JOUR
T1 - Protocol for generating and using human iPSC-derived microglia-containing air-liquid-interface cortical organoid cultures
AU - Cañizares Luna, Marta
AU - Mars, Mayte
AU - Jakobs, Channa E
AU - Huffels, Christiaan F M
AU - Ermakov, Arthur
AU - Hol, Elly M
AU - Pasterkamp, R Jeroen
N1 - Publisher Copyright:
© 2025 The Author(s)
PY - 2025/9/19
Y1 - 2025/9/19
N2 - 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.
AB - 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.
U2 - 10.1016/j.xpro.2025.103915
DO - 10.1016/j.xpro.2025.103915
M3 - Article
C2 - 40601426
SN - 2666-1667
VL - 6
JO - STAR protocols
JF - STAR protocols
IS - 3
M1 - 103915
ER -