title: Small molecule promoted feeder free and adherent differentiation of functional neurons from human embryonic and induced pluripotent stem cells.
abstract: While human embryonic stem (hES) and induced pluripotent stem (hiPS) cells offer exciting prospects in the fields of regenerative medicine and developmental biology, efficient directed differentiation of these cells is still difficult. Neural induction protocols often include suspension culture or co-culture with other cell types, introducing heterogeneity and complicating analysis. In addition, expensive recombinant factors are often used over processes that take weeks to complete, making such experiments financially difficult. We have developed a fully adherent and feeder free neural differentiation protocol using small molecules such as dorsomorphin and common medium supplements. Using this protocol, we obtain >90% of cells developing into neural precursors, as measured by nestin staining. Neurons derived from these precursors are electrophysiologically active. After three weeks of terminal differentiation, we obtain functional neurons which fire high-amplitude action potentials upon depolarization. A subset of neurons also fires repetitive trains. This protocol offers a simpler and less expensive method for investigations involving the differentiation of neural precursors and neurons in culture.
authors.affiliation: Emory University School of Medicine, Department of Anesthesiology, Woodruff Circle, Woodruff Memorial Research Building Suite 617, Atlanta, GA 30322 USA.
authors.forename: Danielle
authors.initials: D
authors.lastname: Drury-Stewart
authors.forename: Mingke
authors.initials: M
authors.lastname: Song
authors.forename: Osama
authors.initials: O
authors.lastname: Mohamad
authors.forename: Shan Ping
authors.initials: SP
authors.lastname: Yu
authors.forename: Ling
authors.initials: L
authors.lastname: Wei
mesh_terms: D018929:Cell Culture Techniques; D002454:Cell Differentiation / Q000187:drug effects*; D017077:Culture Media, Conditioned; D004594:Electrophysiology; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000187:drug effects; D006801:Humans; D007124:Immunoenzyme Techniques; D057026:Induced Pluripotent Stem Cells / Q000166:cytology* / Q000187:drug effects; D009416:Nerve Regeneration; D058953:Neural Stem Cells / Q000166:cytology* / Q000187:drug effects; D009474:Neurons / Q000166:cytology* / Q000187:drug effects; D011720:Pyrazoles / Q000494:pharmacology*; D011743:Pyrimidines / Q000494:pharmacology*
chemical_list: D017077:Culture Media, Conditioned; D011720:Pyrazoles; D011743:Pyrimidines; C516138:dorsomorphin