✍🏼 Yang  Xuejiao, Mu Yuchen, Wu Qianxiang, Zhou Liqiang, Fan Orion R., Lin Quan, Zhu Wenmin, Sun Yi Eve

🏠 Shanghai Institute of Stem Cell Research and Clinical Translation, East Hospital, Tongji University School of Medicine, Shanghai, China

📑 Neural Regeneration Research (2025)

Abstract
The presence or absence of adult neural stem cells in the mammalian forebrain ependyma has been debated for two decades. In this study, we performed single-cell RNA sequencing to investigate the cellular composition of the ependymal surface of the adult mouse forebrain using whole mounts of lateral walls of lateral ventricles. We identified 12 different cell subtypes in the ependymal surface. Immunocytochemical analyses revealed that CD133+ multi-ciliated cells comprised 67.6% of ependymal cells, while the remaining 32.4% were CD133−. CD133+ ependymal cells can be further classified into FOXJ1+/SOX2+/ACTA2+ cells, FLT1+/CD31+/CLDN5+ endothelial-like cells, and PDGFRB+/VTN+/NG2+ pericyte-like cells, as well as endothelial–pericyte-like cells and Foxj1+ endotheliallike cells. CD133− ependymal cells can be further divided into endothelial-like cells, Foxj1+ ependymal cells, Foxj1+ endothelial-like cells, pericyte-like cells, endothelial-pericyte-like cells, VIM+ cells, and cells negative for all of these markers. This comprehensive profiling confirms the heterogeneity of the ependymal surface in the adult mouse forebrain. Debate regarding whether adult ependymal cells contain neural stem cells has arisen because different researchers have examined different populations of ependymal cells. Our study provides a new perspective for investigation of clinical endogenous neural stem cells, ultimately paving the way for stem cell therapies in neurological diseases.