期刊
STEM CELLS
卷 26, 期 4, 页码 886-893出版社
WILEY-BLACKWELL
DOI: 10.1634/stemcells.2007-0620
关键词
stem cell; motor neuron; small molecule; spinal cord
资金
- NICHD NIH HHS [P30 HD03352, P30 HD003352] Funding Source: Medline
- NINDS NIH HHS [R21-NS055261, R01 NS045926, R21 NS055261, P01 NS057778-01A10003, P01 NS057778, R01-NS045926, R01 NS045926-04] Funding Source: Medline
Specification of distinct cell types from human embryonic stem cells (hESCs) is key to the potential application of these naive 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 (similar to 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.
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