Directed differentiation of ventral spinal progenitors and motor neurons from human embryonic stem cells by small molecules.
Specification of distinct cell types from human embryonic stem cells (hESCs) is key to the potential application of these naïve pluripotent cells in regenerative medicine. Determination of the nontarget differentiated populations, which is lacking in the field, is also crucial. Here, we show an efficient differentiation of motor neurons ( approximately 50%) by a simple sequential application of retinoid acid and sonic hedgehog (SHH) in a chemically defined suspension culture. We also discovered that purmorphamine, a small molecule that activates the SHH pathway, could replace SHH for the generation of motor neurons. Immunocytochemical characterization indicated that cells differentiated from hESCs were nearly completely restricted to the ventral spinal progenitor fate (NKX2.2+, Irx3+, and Pax7-), with the exception of motor neurons (HB9+) and their progenitors (Olig2+). Thus, the directed neural differentiation system with small molecules, even without further purification, will facilitate basic and translational studies using human motoneurons at a minimal cost.
Li
Xue-Jun
XJ
Department of Anatomy and the Stem Cell Research Program, Madison, Wisconsin 53705, USA.
Hu
Bao-Yang
BY
Jones
Stefanie A
SA
Zhang
Ying-Sha
YS
Lavaute
Timothy
T
Du
Zhong-Wei
ZW
Zhang
Su-Chun
SC
D000818:Animals; D002454:Cell Differentiation / Q000187:drug effects / Q000502:physiology; D002460:Cell Line; D002478:Cells, Cultured; D019020:Directed Molecular Evolution / Q000379:methods*; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000502:physiology; D053823:Hedgehog Proteins / Q000096:biosynthesis / Q000235:genetics / Q000502:physiology; D000090322:Homeobox Protein Nkx-2.2; D018398:Homeodomain Proteins; D006801:Humans; D051379:Mice; D009025:Morpholines / Q000494:pharmacology; D009046:Motor Neurons / Q000166:cytology* / Q000187:drug effects / Q000502:physiology*; D009687:Nuclear Proteins; D011687:Purines / Q000494:pharmacology; D013116:Spinal Cord / Q000166:cytology / Q000187:drug effects / Q000502:physiology; D013234:Stem Cells / Q000166:cytology / Q000187:drug effects / Q000502:physiology; D014157:Transcription Factors; D014212:Tretinoin / Q000494:pharmacology / Q000502:physiology
D053823:Hedgehog Proteins; D000090322:Homeobox Protein Nkx-2.2; D018398:Homeodomain Proteins; D009025:Morpholines; C000715631:NKX2-2 protein, human; C000715627:Nkx2-2 protein, mouse; D009687:Nuclear Proteins; D011687:Purines; D014157:Transcription Factors; D014212:Tretinoin; C470893:purmorphamine