Chemically induced specification of retinal ganglion cells from human embryonic and induced pluripotent stem cells.
The loss of retinal ganglion cells (RGCs) is the primary pathological change for many retinal degenerative diseases. Although there is currently no effective treatment for this group of diseases, cell transplantation to replace lost RGCs holds great potential. However, for the development of cell replacement therapy, better understanding of the molecular details involved in differentiating stem cells into RGCs is essential. In this study, a novel, stepwise chemical protocol is described for the differentiation of human embryonic stem cells and induced pluripotent stem cells into functional RGCs. Briefly, stem cells were differentiated into neural rosettes, which were then cultured with the Notch inhibitor N-[N-(3,5-difluorophenacetyl)-l-alanyl]-S-phenylglycine t-butyl ester (DAPT). The expression of neural and RGC markers (BRN3A, BRN3B, ATOH7/Math5, γ-synuclein, Islet-1, and THY-1) was examined. Approximately 30% of the cell population obtained expressed the neuronal marker TUJ1 as well the RGC markers. Moreover, the differentiated RGCs generated action potentials and exhibited both spontaneous and evoked excitatory postsynaptic currents, indicating that functional and mature RGCs were generated. In combination, these data demonstrate that a single chemical (DAPT) can induce PAX6/RX-positive stem cells to undergo differentiation into functional RGCs.
Department of Pediatrics, Division of Human Genetics, Department of Anatomy and Neurobiology, Department of Psychiatry and Human Behavior, MitoMed Molecular Diagnostic Laboratory, Department of Pathology, Department of Developmental and Cell Biology, and Department of Ophthalmology, University of California, Irvine, Irvine, California, USA; Division of Human Genetics, Cincinnati Children's Hospital Medical Center, Cincinnati, Ohio, USA.
Hamidreza
H
Riazifar
Yousheng
Y
Jia
Jing
J
Chen
Gary
G
Lynch
Taosheng
T
Huang
D000943:Antigens, Differentiation / Q000096:biosynthesis*; D002454:Cell Differentiation / Q000187:drug effects*; D002460:Cell Line; D004151:Dipeptides / Q000494:pharmacology*; D053595:Embryonic Stem Cells / Q000166:cytology / Q000378:metabolism*; D005260:Female; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology / Q000378:metabolism*; D008297:Male; D012165:Retinal Ganglion Cells / Q000166:cytology / Q000378:metabolism*
D000943:Antigens, Differentiation; D004151:Dipeptides; C419410:N-(N-(3,5-difluorophenacetyl)alanyl)phenylglycine tert-butyl ester
Differentiation; Embryonic stem cells; Induced pluripotent stem cells; Notch signaling; Retinal ganglion cells