Purmorphamine is a tri-substituted purine derivative that activates the Hedgehog pathway by directly binding to and activating the Hedgehog receptor Smoothened (EC₅₀ = 1 µM). (Sinha and Chen)
DIFFERENTIATION
· Promotes differentiation of ventral spinal progenitors and motor neurons from human pluripotent stem cells (Hu and Zhang, Karumbayaram et al., Li et al.).
· Promotes differentiation of osteoblasts from human and mouse mesenchymal cells (Beloti et al., Wu et al. 2002, Wu et al. 2004).
· Inhibits differentiation and maturation of adipocytes from human mesenchymal cells (Fontaine et al.).
DIFFERENTIATION
· Promotes differentiation of ventral spinal progenitors and motor neurons from human pluripotent stem cells (Hu and Zhang, Karumbayaram et al., Li et al.).
· Promotes differentiation of osteoblasts from human and mouse mesenchymal cells (Beloti et al., Wu et al. 2002, Wu et al. 2004).
· Inhibits differentiation and maturation of adipocytes from human mesenchymal cells (Fontaine et al.).
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Protocols and Documentation
Find supporting information and directions for use in the Product Information Sheet or explore additional protocols below.
Applications
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Resources and Publications
Educational Materials (4)
Publications (8)
Differentiation of spinal motor neurons from pluripotent human stem cells. Nature protocols 2009 JAN
Abstract
We have devised a reproducible protocol by which human embryonic stem cells (hESCs) or inducible pluripotent stem cells (iPSCs) are efficiently differentiated to functional spinal motor neurons. This protocol comprises four major steps. Pluripotent stem cells are induced to form neuroepithelial (NE) cells that form neural tube-like rosettes in the absence of morphogens in the first 2 weeks. The NE cells are then specified to OLIG2-expressing motoneuron progenitors in the presence of retinoic acid (RA) and sonic hedgehog (SHH) or purmorphamine in the next 2 weeks. These progenitor cells further generate post-mitotic, HB9-expressing motoneurons at the 5th week and mature to functional motor neurons thereafter. It typically takes 5 weeks to generate the post-mitotic motoneurons and 8-10 weeks for the production of functional mature motoneurons. In comparison with other methods, our protocol does not use feeder cells, has a minimum dependence on proteins (purmorphamine replacing SHH), has controllable adherent selection and is adaptable for scalable suspension culture.
Directed differentiation of human-induced pluripotent stem cells generates active motor neurons. Stem cells (Dayton, Ohio) 2009 APR
Abstract
The potential for directed differentiation of human-induced pluripotent stem (iPS) cells to functional postmitotic neuronal phenotypes is unknown. Following methods shown to be effective at generating motor neurons from human embryonic stem cells (hESCs), we found that once specified to a neural lineage, human iPS cells could be differentiated to form motor neurons with a similar efficiency as hESCs. Human iPS-derived cells appeared to follow a normal developmental progression associated with motor neuron formation and possessed prototypical electrophysiological properties. This is the first demonstration that human iPS-derived cells are able to generate electrically active motor neurons. These findings demonstrate the feasibility of using iPS-derived motor neuron progenitors and motor neurons in regenerative medicine applications and in vitro modeling of motor neuron diseases.
Directed differentiation of ventral spinal progenitors and motor neurons from human embryonic stem cells by small molecules. Stem cells (Dayton, Ohio) 2008 APR
Abstract
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.
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