title: Analysis of in vitro and in vivo characteristics of human embryonic stem cell-derived neural precursors.
abstract: 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.
authors.affiliation: Institute of Experimental Medicine, Academy of Sciences of the Czech Republic, Prague, Czech Republic.
authors.forename: Nataliya
authors.initials: N
authors.lastname: Kozubenko
authors.forename: Karolina
authors.initials: K
authors.lastname: Turnovcova
authors.forename: Miroslava
authors.initials: M
authors.lastname: Kapcalova
authors.forename: Olena
authors.initials: O
authors.lastname: Butenko
authors.forename: Miroslava
authors.initials: M
authors.lastname: Anderova
authors.forename: Vendula
authors.initials: V
authors.lastname: Rusnakova
authors.forename: Mikael
authors.initials: M
authors.lastname: Kubista
authors.forename: Ales
authors.initials: A
authors.lastname: Hampl
authors.forename: Pavla
authors.initials: P
authors.lastname: Jendelova
authors.forename: Eva
authors.initials: E
authors.lastname: Sykova
mesh_terms: D000818:Animals; D015415:Biomarkers / Q000378:metabolism*; D001921:Brain / Q000378:metabolism / Q000473:pathology; D002454:Cell Differentiation; D002460:Cell Line; D019070:Cell Lineage; D064987:Cell- and Tissue-Based Therapy; D053595:Embryonic Stem Cells / Q000166:cytology*; D005434:Flow Cytometry; D006801:Humans; D007150:Immunohistochemistry; D009474:Neurons / Q000166:cytology / Q000378:metabolism / Q000637:transplantation*; D010641:Phenotype; D051381:Rats
chemical_list: D015415:Biomarkers