Glutamatergic neuronal differentiation of mouse embryonic stem cells after transient expression of neurogenin 1 and treatment with BDNF and GDNF: in vitro and in vivo studies.
Differentiation of the pluripotent neuroepithelium into neurons and glia is accomplished by the interaction of growth factors and cell-type restricted transcription factors. One approach to obtaining a particular neuronal phenotype is by recapitulating the expression of these factors in embryonic stem (ES) cells. Toward the eventual goal of auditory nerve replacement, the aim of the current investigation was to generate auditory nerve-like glutamatergic neurons from ES cells. Transient expression of Neurog1 promoted widespread neuronal differentiation in vitro; when supplemented with brain-derived neurotrophic factor (BDNF) and glial cell line-derived neurotrophic factor (GDNF), 75% of ES cell-derived neurons attained a glutamatergic phenotype after 5 d in vitro. Mouse ES cells were also placed into deafened guinea pig cochleae and Neurog1 expression was induced for 48 h followed by 26 d of BDNF/GDNF infusion. In vivo differentiation resulted in 50-75% of ES cells bearing markers of early neurons, and a majority of these cells had a glutamatergic phenotype. This is the first study to report a high percentage of ES cell differentiation into a glutamatergic phenotype and sets the stage for cell replacement of auditory nerve.
Reyes
Jeannie H
JH
Kresge Hearing Research Institute, Department of Otolaryngology, University of Michigan, Ann Arbor, Michigan 48109-0506, USA.
O'Shea
K Sue
KS
Wys
Noel L
NL
Velkey
J Matthew
JM
Prieskorn
Diane M
DM
Wesolowski
Karolina
K
Miller
Josef M
JM
Altschuler
Richard A
RA
D000818:Animals; D051792:Basic Helix-Loop-Helix Proteins / Q000235:genetics / Q000378:metabolism*; D015415:Biomarkers / Q000032:analysis / Q000378:metabolism; D019208:Brain-Derived Neurotrophic Factor / Q000378:metabolism / Q000494:pharmacology; D002454:Cell Differentiation / Q000187:drug effects / Q000502:physiology; D002460:Cell Line; D002478:Cells, Cultured; D003056:Cochlear Nerve / Q000187:drug effects / Q000196:embryology / Q000378:metabolism; D003638:Deafness / Q000139:chemically induced / Q000378:metabolism / Q000601:surgery; D004318:Doxycycline / Q000494:pharmacology; D053595:Embryonic Stem Cells / Q000187:drug effects / Q000378:metabolism / Q000637:transplantation*; D051100:Glial Cell Line-Derived Neurotrophic Factor / Q000378:metabolism / Q000494:pharmacology; D018698:Glutamic Acid / Q000378:metabolism*; D006168:Guinea Pigs; D006801:Humans; D051379:Mice; D009414:Nerve Growth Factors / Q000378:metabolism / Q000494:pharmacology*; D009419:Nerve Tissue Proteins / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D055495:Neurogenesis / Q000187:drug effects / Q000502:physiology*; D010641:Phenotype; D011984:Sensory Receptor Cells / Q000187:drug effects / Q000378:metabolism; D033581:Stem Cell Transplantation / Q000379:methods*
D051792:Basic Helix-Loop-Helix Proteins; D015415:Biomarkers; D019208:Brain-Derived Neurotrophic Factor; D004318:Doxycycline; D051100:Glial Cell Line-Derived Neurotrophic Factor; D018698:Glutamic Acid; D009414:Nerve Growth Factors; D009419:Nerve Tissue Proteins; C486336:Neurog1 protein, mouse