Differentiation of human embryonic stem cells to regional specific neural precursors in chemically defined medium conditions.
BACKGROUND
Human 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.


RESULTS
The 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
These 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.
Erceg
Slaven
S
Cellular Reprogramming Laboratory, Centro de Investigación Príncipe Felipe (CIPF), Valencia, Spain.
Laínez
Sergio
S
Ronaghi
Mohammad
M
Stojkovic
Petra
P
Pérez-Aragó
Maria Amparo
MA
Moreno-Manzano
Victoria
V
Moreno-Palanques
Rubén
R
Planells-Cases
Rosa
R
Stojkovic
Miodrag
M
D018929:Cell Culture Techniques / Q000379:methods; D002454:Cell Differentiation / Q000502:physiology*; D002460:Cell Line; D003470:Culture Media; D004594:Electrophysiology; D047108:Embryonic Development / Q000502:physiology*; D053595:Embryonic Stem Cells / Q000166:cytology*; D006801:Humans; D007150:Immunohistochemistry; D009474:Neurons / Q000166:cytology* / Q000502:physiology*; D009928:Organ Specificity; D018408:Patch-Clamp Techniques; D012313:RNA / Q000235:genetics / Q000302:isolation & purification; D020133:Reverse Transcriptase Polymerase Chain Reaction
D003470:Culture Media; D012313:RNA