Quick and robust method for the generation of human iPSC-derived choroid plexus organoids

Abstract

The choroid plexus (ChP) is a key brain structure responsible for cerebrospinal fluid (CSF) production and forms a selective barrier that regulates brain homeostasis and immune surveillance. In vitro models of ChP are essential for studying CSF dynamics, viral entry, neuroinflammation, and CNS drug transport, yet current organoid protocols remain complex, slow, and difficult to reproduce. Here, we report a quick and robust method for the generation of human iPSC-derived ChP organoids that is xeno-free/ serum-free, scalable, and reproducible. Early GSK3{beta} inhibition and transient WNT modulation guide organoids toward cystic ChP-enriched structures, confirmed by ventricle-like morphology and expression of canonical markers (TTR, ZO-1). This minimal workflow enables rapid production of functional ChP-like organoids suitable for CSF physiology studies, barrier modeling, and translational neuroscience, providing an accessible platform for both basic and applied research.