iPSC-Derived Human Motor Neurons Starter Kit (African-American, Male Line)
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Motor neurons (MN) are a specialized type of neurons originating from the spinal cord that are responsible for integrating signals from the brain and the muscles to control and coordinate voluntary and involuntary contraction of muscles and movement. Motor neuron degeneration or dysfunction has been implicated in serious and debilitating diseases such as spinal muscular atrophy (SMA), amyotrophic lateral scleoris (ALS), post-polio syndrome (PPS) and others. iPSC-derived motor neurons provide a convenient, consistent, reliable and physiologically relevant source of cell line mdels to model diseases and to test novel therapies.
Applied StemCell has developed an efficient integration-free, small molecule-based method to differentiate high-quality motor neuron cells from human iPSCs. Our proprietary differentiation protocol involves both the activation of the Wnt pathway, SHH signaling, and the inhibition of the activin-nodal and BMP signaling. The differentiated motor neurons recapitulate the phenotype and functional parameters of primary and in vivo motor neurons.
We provide motor neurons differentiated from an integration-free, control human iPSC line (ASE-9211), reprogrammed from fibroblasts of an African-American male donor. These high-purity (≥90%) cells show distinct neurite outgrowth in 2-7 days after thaw (Figure 1) and express late-stage motor neuron precursor biomarkers, HB9 and ChAT at day 2 after thaw (Figure 2) , and mature motor neuron biomarkers, Tuj1 and MAP2 at day 5 (Figure 3). The cells are provided as cryopreserved, late-stage precursors that mature in 5 days after recovery.
To harness the full potential of our motor neurons, we also provide optimized, serum-free Motor Neuron Culture Media (ASE-9701MM) that supports robust maintenance and functionality of the motor neurons in culture.
These iPSC-differentiated motor neurons can be used as control lines to compare phenotype and functionality of patient-derived and genome edited iPSC-derived motor neurons, for co-culture models with other neurons, glia and skeletal muscle cells, as well as for neurotoxicity and drug screening.
Characterization of the ASE-9701 Motor Neurons
Figure 1. Recovery of cryopreserved ASE-9701 iPSC-derived Motor Neurons. Cryopreserved motor neurons, differentiated from Applied StemCell’s control iPSC line, ASE-9211 were recovered in motor neuron culture media and undergo fast maturation in 5 days. Images at 20x magnification.
Figure 2. Immunostaining of ASE-9701 iPSC-derived Motor Neurons for motor neuron biomarkers. Cryopreserved motor neurons, differentiated from Applied StemCell’s control iPSC line, ASE-9211 were recovered in motor neuron culture media The cells were stained for motor neuron late-stage precursor markers, HB9 (green) and TUJ1 (red) at day2,. DAPI: nuclear counterstain (blue). Images at 20x magnification.
Figure 3. Immunostaining of ASE-9701 iPSC-derived Motor Neurons for motor neuron biomarkers. Cryopreserved motor neurons, differentiated from Applied StemCell’s control iPSC line, ASE-9211 were recovered in motor neuron culture media The cells were stained for mature motor neuron markers, CHAT (green) and pan-neuronal marker MAP2 (red) at day5. DAPI: nuclear counterstain (blue). Images at 20x magnification.