Journal
CELL TRANSPLANTATION
Volume 19, Issue 4, Pages 471-486Publisher
SAGE PUBLICATIONS INC
DOI: 10.3727/096368909X484707
Keywords
Human embryonic stem cells (hESCs); Neural differentiation; Fluorescent-activated cell sorting; RT-PCR; Transplantation
Funding
- Academy of Sciences of the Czech Republic [AV0Z50390703, AVO250520701]
- Ministry of Education, Youth and Sports of the Czech Republic [1M0538, LC554]
- Academy of Science [IAA500520809]
- Grant Agency of the Czech Republic [305/09/0717, 309/08/H079]
- EC [FP6, LSHB-CT-2006-037328, LSHM-CT-2005-019063]
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During the last decade, much progress has been made in developing protocols for the differentiation of human embryonic stem cells (hESCs) into a neural phenotype. The appropriate agent for cell therapy is neural precursors (NPs). Here, we demonstrate the derivation of highly enriched and expandable populations of proliferating NPs from the CCTL14 line of hESCs. These NPs could differentiate in vitro into functionally active neurons, as confirmed by immunohistochemical staining and electrophysiological analysis. Neural cells differentiated in vitro from hESCs exhibit broad cellular heterogeneity with respect to developmental stage and lineage specification. To analyze the population of the derived NPs, we used fluorescence-activated cell sorting (FACS) and characterized the expression of several pluripotent and neural markers, such as Nanog, SSEA-4, SSEA-1, TRA-1-60, CD24, CD133, CD56 (NCAM), beta-III-tubulin, NF70, nestin, CD271 (NGFR), CD29, CD73, and CD105 during long-term propagation. The analyzed cells were used for transplantation into the injured rodent brain; the tumorigenicity of the transplanted cells was apparently eliminated following long-term culture. These results complete the characterization of the CCTL14 line of hESCs and provide a framework for developing cell selection strategies for neural cell-based therapies.
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