{"doi":"10.1016/j.xpro.2025.104142","title":"Protocol for purifying primitive neural progenitors by fractional passaging to generate astroglia-enriched cerebral organoid","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 • Instructions for differentiating hPSCs into pNPC cultures • Steps for fractional passaging yield highly pure pNPCs consistently across cell lines • Procedures for generating astroglia-enriched cerebral organoids Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. 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.","journal":"STAR Protocols","year":2025,"id":578072,"datarank":0.0,"base_score":0.0,"endowment":0.0,"self_citation_contribution":0.0,"citation_network_contribution":0.0,"self_endowment_contribution":0.0,"citer_contribution":0.0,"corpus_percentile":null,"corpus_rank":null,"citation_count":0,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9538,"is_data_producer":false,"deposit_databanks":null,"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2025-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":1328472,"name":"Rui Dang","orcid":"0000-0003-4572-6879","position":1,"is_corresponding":false},{"id":623006,"name":"Mengmeng Jin","orcid":"0000-0002-2749-8000","position":2,"is_corresponding":false},{"id":245303,"name":"Peng Jiang","orcid":"0000-0002-2650-3082","position":3,"is_corresponding":false},{"id":1382135,"name":"Kushal Aluru","orcid":null,"position":0,"is_corresponding":true}],"reference_count":20,"raw_metadata":null,"created_at":"2026-07-19T02:58:20.638044Z","pmid":"41091601","pmcid":null,"fwci":null,"citation_percentile":null,"influential_citations":0,"oa_status":null,"license":null,"views":0,"total_file_size_bytes":0,"version_count":0,"fair_f":null,"fair_a":null,"fair_i":null,"fair_r":null,"fair_zscore":null,"fair_rationale":null,"fair_model":null,"fair_agent_version":null,"fair_fulltext_source":null,"fair_has_llm":null,"fair_computed_at":null,"clinical_trials":[],"software_tools":[],"db_accessions":[],"linked_datasets":[],"topics":[]}