Cell-Surface Marker Signatures for the Isolation of Neural Stem Cells, Glia and Neurons Derived from Human Pluripotent Stem Cells

PLoS One

Jason G. Vidal¹  Christian T. Carson¹  Lawrence S. B. Goldstein¹  Jessica Flippin²  Jeanne Elia²  Yangling Mu²  Fred H. Gage²  Mike P. Hefferan²  Nil Emre³  Martin Marsala³  Jody Martin⁴  Mason A. Israel⁵  Rosanto I. Paramban⁵  Silvia Marsala⁵  Shauna H. Yuan⁵  Rhiannon L. Killian⁶ 

[1] Anesthesiology Research Laboratory, Department of Anesthesiology, University of California San Diego, La Jolla, California, United States of America;BD Biosciences, La Jolla, California, United States of America;Biomedical Sciences Graduate Program, University of California San Diego, La Jolla, California, United States of America;Department of Neurosciences, School of Medicine, University of California San Diego, La Jolla, California, United States of America;Howard Hughes Medical Institute and Department of Cellular and Molecular Medicine, School of Medicine, University of California San Diego, La Jolla, California, United States of America;Laboratory of Genetics, The Salk Institute for Biological Studies, La Jolla, California, United States of America

关键词: Neuronal differentiation;    Neurons;    Cell staining;    DAPI staining;    Astrocytes;    Contaminants;    Selection markers;    Neural stem cells;   

DOI  :  10.1371/journal.pone.0017540

学科分类：医学（综合）

来源: Public Library of Science

【 摘 要 】

Background Neural induction of human pluripotent stem cells often yields heterogeneous cell populations that can hamper quantitative and comparative analyses. There is a need for improved differentiation and enrichment procedures that generate highly pure populations of neural stem cells (NSC), glia and neurons. One way to address this problem is to identify cell-surface signatures that enable the isolation of these cell types from heterogeneous cell populations by fluorescence activated cell sorting (FACS). Methodology/Principal Findings We performed an unbiased FACS- and image-based immunophenotyping analysis using 190 antibodies to cell surface markers on naïve human embryonic stem cells (hESC) and cell derivatives from neural differentiation cultures. From this analysis we identified prospective cell surface signatures for the isolation of NSC, glia and neurons. We isolated a population of NSC that was CD184+/CD271−/CD44−/CD24+ from neural induction cultures of hESC and human induced pluripotent stem cells (hiPSC). Sorted NSC could be propagated for many passages and could differentiate to mixed cultures of neurons and glia in vitro and in vivo. A population of neurons that was CD184−/CD44−/CD15LOW/CD24+ and a population of glia that was CD184+/CD44+ were subsequently purified from cultures of differentiating NSC. Purified neurons were viable, expressed mature and subtype-specific neuronal markers, and could fire action potentials. Purified glia were mitotic and could mature to GFAP-expressing astrocytes in vitro and in vivo. Conclusions/Significance These findings illustrate the utility of immunophenotyping screens for the identification of cell surface signatures of neural cells derived from human pluripotent stem cells. These signatures can be used for isolating highly pure populations of viable NSC, glia and neurons by FACS. The methods described here will enable downstream studies that require consistent and defined neural cell populations.

【 授权许可】

CC BY