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  3. Protocol for purifying primitive neural progenitors by fractional passaging to generate astroglia-enriched cerebral organoid

Protocol for purifying primitive neural progenitors by fractional passaging to generate astroglia-enriched cerebral organoid

  • STAR Protoc. 2025 Oct 13;6(4):104142. doi: 10.1016/j.xpro.2025.104142.
Kushal Aluru 1,  Rui Dang 2,  Mengmeng Jin 3,  Peng Jiang 4
Affiliations

Affiliations

  • 1 Department of Cell Biology and Neuroscience, Rutgers University-New Brunswick, Piscataway, NJ 08854, USA. Electronic address: kushal.aluru@rutgers.edu.
  • 2 Department of Cell Biology and Neuroscience, Rutgers University-New Brunswick, Piscataway, NJ 08854, USA. Electronic address: rui.dang@rutgers.edu.
  • 3 Department of Cell Biology and Neuroscience, Rutgers University-New Brunswick, Piscataway, NJ 08854, USA. Electronic address: mengmeng.jin@rutgers.edu.
  • 4 Department of Cell Biology and Neuroscience, Rutgers University-New Brunswick, Piscataway, NJ 08854, USA. Electronic address: peng.jiang@rutgers.edu.
PMID: 41091601 DOI: 10.1016/j.xpro.2025.104142
Abstract

Human pluripotent stem cell (hPSC)-derived primitive neural progenitor cells (pNPCs) and cerebral organoids provide a powerful platform for studying human brain development. Here, we present a fractional passaging protocol that yields nearly pure PAX6+/SOX2+ pNPCs. We describe steps for neuroepithelial differentiation, purification, and production of astroglia-enriched organoids. We then detail procedures for downstream analysis. The organoids contain over 15% astrocytes alongside maturing neurons, providing a scalable and reproducible system to model human glial-neuronal interactions in development and disease. For complete details on the use and execution of this protocol, please refer to Dang et al.1.

Keywords

Neuroscience; Organoids; Stem Cells.

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