Olga Vafaeva, Poommaree Namchaiw, Karl Murray, Elva Diaz, Hwai-Jong Cheng
{"title":"Protocol for culturing neurospheres from progenitor cells in the dentate gyrus of aged mouse hippocampus.","authors":"Olga Vafaeva, Poommaree Namchaiw, Karl Murray, Elva Diaz, Hwai-Jong Cheng","doi":"10.1016/j.xpro.2025.103692","DOIUrl":null,"url":null,"abstract":"<p><p>The neurosphere assay is the gold standard for assessing the proliferative and differentiation capacities of neural progenitor cells (NPCs). Here, we present a protocol for isolating, propagating, and maintaining hippocampal neurospheres from adult and aged mice and differentiating cultured NPCs into neurons and astrocytes. We describe steps for establishing a heterochronic co-culture of neurosphere-derived cells with primary neurons. Using neurospheres from old animals enables investigation of the effects of aging on the development and differentiation of newborn neurons.</p>","PeriodicalId":34214,"journal":{"name":"STAR Protocols","volume":"6 1","pages":"103692"},"PeriodicalIF":1.3000,"publicationDate":"2025-03-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"STAR Protocols","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.1016/j.xpro.2025.103692","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q4","JCRName":"BIOCHEMICAL RESEARCH METHODS","Score":null,"Total":0}
引用次数: 0
Abstract
The neurosphere assay is the gold standard for assessing the proliferative and differentiation capacities of neural progenitor cells (NPCs). Here, we present a protocol for isolating, propagating, and maintaining hippocampal neurospheres from adult and aged mice and differentiating cultured NPCs into neurons and astrocytes. We describe steps for establishing a heterochronic co-culture of neurosphere-derived cells with primary neurons. Using neurospheres from old animals enables investigation of the effects of aging on the development and differentiation of newborn neurons.