Open AccessMarkers and Methods for Cell Sorting of Human Embryonic Stem Cell‐Derived Neural Cell Populations
Author(s) -
Pruszak Jan,
Sonntag KaiChristian,
Aung Moe Hein,
SanchezPernaute Rosario,
Isacson Ole
Publication year - 2007
Publication title -
stem cells
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.159
H-Index - 229
eISSN - 1549-4918
pISSN - 1066-5099
DOI - 10.1634/stemcells.2006-0744
Subject(s) - biology , embryonic stem cell , cell sorting , neural cell adhesion molecule , microbiology and biotechnology , neural development , cd146 , cellular differentiation , neural stem cell , neural cell , stem cell , cell , flow cytometry , cell adhesion , genetics , gene , cd34
Abstract Neural cells differentiated in vitro from human embryonic stem cells (hESC) exhibit broad cellular heterogeneity with respect to developmental stage and lineage specification. Here, we describe standard conditions for the use and discovery of markers for analysis and cell selection of hESC undergoing neuronal differentiation. To generate better‐defined cell populations, we established a working protocol for sorting heterogeneous hESC‐derived neural cell populations by fluorescence‐activated cell sorting (FACS). Using genetically labeled synapsin‐green fluorescent protein‐positive hESC‐derived neurons as a proof of principle, we enriched viable differentiated neurons by FACS. Cell sorting methodology using surface markers was developed, and a comprehensive profiling of surface antigens was obtained for immature embryonic stem cell types (such as stage‐specific embryonic antigen [SSEA]‐3, ‐4, TRA‐1‐81, TRA‐1‐60), neural stem and precursor cells (such as CD133, SSEA‐1 [CD15], A2B5, forebrain surface embryonic antigen‐1, CD29, CD146, p75 [CD271]), and differentiated neurons (such as CD24 or neural cell adhesion molecule [NCAM; CD56]). At later stages of neural differentiation, NCAM (CD56) was used to isolate hESC‐derived neurons by FACS. Such FACS‐sorted hESC‐derived neurons survived in vivo after transplantation into rodent brain. These results and concepts provide (a) a feasible approach for experimental cell sorting of differentiated neurons, (b) an initial survey of surface antigens present during neural differentiation of hESC, and (c) a framework for developing cell selection strategies for neural cell‐based therapies. Disclosure of potential conflicts of interest is found at the end of this article.