title: Cell-surface marker signatures for the isolation of neural stem cells, glia and neurons derived from human pluripotent stem cells.
abstract: 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).
  
  
  RESULTS
  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(-)/CD15(LOW)/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
  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.
authors.affiliation: 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.
authors.forename: Shauna H
authors.initials: SH
authors.lastname: Yuan
authors.forename: Jody
authors.initials: J
authors.lastname: Martin
authors.forename: Jeanne
authors.initials: J
authors.lastname: Elia
authors.forename: Jessica
authors.initials: J
authors.lastname: Flippin
authors.forename: Rosanto I
authors.initials: RI
authors.lastname: Paramban
authors.forename: Mike P
authors.initials: MP
authors.lastname: Hefferan
authors.forename: Jason G
authors.initials: JG
authors.lastname: Vidal
authors.forename: Yangling
authors.initials: Y
authors.lastname: Mu
authors.forename: Rhiannon L
authors.initials: RL
authors.lastname: Killian
authors.forename: Mason A
authors.initials: MA
authors.lastname: Israel
authors.forename: Nil
authors.initials: N
authors.lastname: Emre
authors.forename: Silvia
authors.initials: S
authors.lastname: Marsala
authors.forename: Martin
authors.initials: M
authors.lastname: Marsala
authors.forename: Fred H
authors.initials: FH
authors.lastname: Gage
authors.forename: Lawrence S B
authors.initials: LS
authors.lastname: Goldstein
authors.forename: Christian T
authors.initials: CT
authors.lastname: Carson
mesh_terms: D000818:Animals; D000906:Antibodies / Q000378:metabolism; D015703:Antigens, CD / Q000378:metabolism; D015415:Biomarkers / Q000378:metabolism; D002454:Cell Differentiation; D002462:Cell Membrane / Q000378:metabolism*; D049109:Cell Proliferation; D002469:Cell Separation / Q000379:methods*; D002470:Cell Survival; D002478:Cells, Cultured; D006801:Humans; D051379:Mice; D008954:Models, Biological; D058953:Neural Stem Cells / Q000166:cytology* / Q000378:metabolism / Q000637:transplantation; D009457:Neuroglia / Q000166:cytology* / Q000378:metabolism; D009474:Neurons / Q000166:cytology* / Q000378:metabolism; D010641:Phenotype; D039904:Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D013116:Spinal Cord / Q000473:pathology
chemical_list: D000906:Antibodies; D015703:Antigens, CD; D015415:Biomarkers