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Abstract
BackgroundHuman embryonic stem cells (hESC) provide a unique model to study early events in human development. The hESC-derived cells can potentially be used to replace or restore different tissues including neuronal that have been damaged by disease or injury.Methodology and Principal FindingsThe cells of two different hESC lines were converted to neural rosettes using adherent and chemically defined conditions. The progenitor cells were exposed to retinoic acid (RA) or to human recombinant basic fibroblast growth factor (bFGF) in the late phase of the rosette formation. Exposing the progenitor cells to RA suppressed differentiation to rostral forebrain dopamine neural lineage and promoted that of spinal neural tissue including motor neurons. The functional characteristics of these differentiated neuronal precursors under both, rostral (bFGF) and caudalizing (RA) signals were confirmed by patch clamp analysis.Conclusions/SignificanceThese findings suggest that our differentiation protocol has the capacity to generate region-specific and electrophysiologically active neurons under in vitro conditions without embryoid body formation, co-culture with stromal cells and without presence of cells of mesodermal or endodermal lineages.
Highlights
Human embryonic stem cells are pluripotent cells that can be propagated in vitro for a long period and represent a theoretically inexhaustible source of precursor cells that could be differentiated into any cell type to study human development or treat debilitating diseases [1,2,3]
We have demonstrated that neural progenitors can be efficiently generated in a one step approach using feeder-free and chemically defined conditions but without formation of embryoid bodies (EBs)
Exposure of Human embryonic stem cells (hESC) to different human extracellular matrix (ECM) substrates leads to the generation of highly enriched neural progenitors without presence of other cells with mesodermal or endodermal characteristics
Summary
Human embryonic stem cells (hESC) are pluripotent cells that can be propagated in vitro for a long period and represent a theoretically inexhaustible source of precursor cells that could be differentiated into any cell type to study human development or treat debilitating diseases [1,2,3]. Derivation of neural progenitors from hESC holds promise to investigate human neurogenesis, to study the development of the central nervous system (CNS) and for potential cell therapy applications to treat Parkinson’s disease or spinal cord injury [4]. At this time, hESC differentiation towards neural lineages has several concerns. The current protocols used to induce neural conversion of hESC include the presence of stromal cell lines (PA6 or MS5), Matrigel or conditioned medium including a multistep procedure which involves formation of embryoid bodies (EBs) [3,5,6,7,8,9,10,11,12,13]. The hESC-derived cells can potentially be used to replace or restore different tissues including neuronal that have been damaged by disease or injury
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