Motor Neurons Differentiation Service

Based on the advanced STEMOD™ platform we have established, Creative Biolabs is proud to offer high-quality motor neurons (MNs) differentiation service for our global customers. The service offered by Creative Biolabs will contribute greatly to the success of your project.

Introduction to MNs

MNs are neuronal cells located in the central nervous system (CNS) for controlling various downstream targets. MNs have the most common type of body plan for a nerve cell-they are multipolar, each with one axon and several dendrites. Spinal MNs located in the ventral horn of the spinal cord are responsible for controlling effector muscles in the periphery and control effector muscles in the periphery. Spinal MNs connect to muscles, glands, and organs throughout the body and form neuronal circuitry. These neurons transmit impulses and control all of our muscle movements.

The generation of MNs appears to involve several developmental steps. Ectodermal cells acquire an initial rostral neural character through the regulation of BMP, FGF, and Wnt signaling. These rostral neural progenitors acquire a spinal positional identity to scandalize signals that include retinoic acid (RA). Subsequently, spinal progenitor cells acquire an MN progenitor identity in response to the ventralizing action of Sonic hedgehog (Shh).

• Classification of MNs

MNs are divided into two categories according to the locations: (1) upper MNs that originate from the cerebral cortex, and (2) lower MNs that are located in the brainstem and spinal cord. Lower MNs travel from the spinal cord to muscle, whereas upper MNs travel between the brain and spinal cord. Lower MNs are alpha MNs, beta MN, and gamma MNs, including branchial MNs, visceral MNs, and somatic MNs.

Fig.1 Characteristics of alpha and gamma MNs.¹

• Functions of MNs

MNs integrate signals from the brain and the sensory systems to control all voluntary and involuntary movements and parts of the autonomic nervous system. MNs can form the efferent division of the PNS. There are approximately 500,000 MNs carrying information from the CNS to peripheral effectors in peripheral tissues and organ systems. Efferent fibers are the axons of MNs that carry information away from the CNS. A single MN may innervate many muscle fibers, and a muscle fiber can undergo most action potentials in the time taken for a single muscle twitch. Innervation occurs at a neuromuscular junction, and twitches can become superimposed.

• Diseases Related to MNs

MNs target devastating diseases, including amyotrophic lateral sclerosis (ALS), spinal muscular atrophy, multiple sclerosis, progressive muscular atrophy, and spinal cord injuries. Evidence indicated that in the early stages of ALS, the nerve terminals and MN junctions are partially degraded. Selective vulnerability of MNs likely arises from several mechanisms, including protein misfolding, mitochondrial dysfunction, oxidative damage, defective axonal transport, excitotoxicity, insufficient growth factor signaling, and inflammation. Nonneuronal neighboring cells' damage enhances damage within MNs via an inflammatory response that accelerates disease progression.

Fig.2 Schematic of the evolution of MN degeneration and glial activation during SOD1 mutant-initiated ALS disease.²

MNs Differentiation Service

With the advent of induced stem cells offering patient-specific treatment hopes, stem cells have been shown to generate MNs in vitro under developmental cues. Stem cells-derived MNs are a valuable tool for many MN studies. Stem cells-derived MNs make excellent candidates for drug screening, particularly in human-derived cells where in vivo tests are inappropriate, and iPSCs can be generated from patients. As a leading custom service provider in neuroscience ex vivo models, Creative Biolabs is devoted to solving numerous challenging projects. With over a decade of extensive experience in providing custom neural differentiation services, our scientists contributed to establishing an advanced STEMOD™ platform to meet every specific requirement from our customers.

Based on our most advanced techniques and years of experience, Creative Biolabs is dedicated to assisting our clients with the most satisfactory stem cell-based ex vivo model-related solutions. If you are interested in learning more about our capacity, please do not hesitate to contact us.

• Our service starts with an initial consultation where we analyze client needs and determine the optimal approach for their specific project.
• After the consultation, we embark on the differentiation process, which includes several stages. Initially, we start with the induction of pluripotent stem cells into the neural lineage, then proceed with the patterning of these neural progenitor cells into spinal cord motor neurons. Once the differentiation is successful, further maturation is induced to achieve fully functioning motor neurons.
• We routinely perform validation checks that typically include marker gene expression, immunofluorescence, and live/dead assays. This ensures optimal cell health, purity, and survival rates, leading to reliable, high-quality motor neurons.
• In addition, we provide comprehensive after-service support to all our customers. This includes providing advice about cell culture, troubleshooting, and protocols for downstream applications.

To conclude, we maintain the utmost commitment to ensure consistent and reproducible results every single time deserving the trust that our clients put in our motor neuron differentiation service. By rendering this service and related services, we aim to accelerate research and drug discovery in neurodegenerative diseases, including but not limited to:

Published Data

He Jax Xu et al. established human spinal cord neural progenitor cells (hSCNPCs) from hPSC-differentiated neuro mesodermal progenitor cells (NMPs) and demonstrated that hSCNPCs can be continuously expanded to 40 generations. The hSCNPCs can efficiently and rapidly differentiate into posterior spinal cord motor neurons. Functional maturity has been examined in detail.

To comprehensively study spinal motor neuron maturation, they utilized the CMOS-based HD-MEA platform, a high-throughput, high-resolution system containing 26,400 electrodes and 1024 simultaneous recording channels, to examine spontaneous neural activity during spinal motor neuron differentiation. As shown, these results indicate that hSCNPC can further differentiate into homogeneous spinal motor neurons and exhibit post-spinal properties. All of the above parameters increased steadily with the progression of differentiation, suggesting a gradual maturation of the electrophysiology of spinal motor neurons.

Fig. 3 Functional characterization of hSCNPCs direct differentiated posterior spinal motor neurons.³

Applications

Our motor neuron differentiation service offers a specialized solution for researchers and pharmaceutical companies involved in neurological studies or drug discovery. Here's a detailed description of its application.

• Basic Research - The motor neurons produced through our differentiation service can be invaluable tools for studying neuronal development, function, and communication in healthy and pathological conditions.
• Disease Modeling - Motor neuron diseases like ALS, SMA, and other neuropathies can be modeled in a dish, which provides better insight into the development and progression of these diseases and potential therapeutic targets.
• Drug Discovery and Testing - These cells can be used as in vitro models to test the effectiveness, neurotoxicity and side effects of new drugs.
• Cell Therapies - For conditions like ALS or SMA where specific populations of motor neurons degenerate, producing mass quantities of identical motor neurons may provide a source of cells for transplantation therapies.

FAQs

• Q: Can the differentiated motor neurons be immediately used for experiments or do they require additional processing?
  A: Differentiated motor neurons can be used instantly for some experiments. However, additional processing may be required for specific experimental setups. This might include further maturation stages, culture with other cell types, or plating in formats suitable for your assays.
• Q: Are there any specific requirements or recommendations for the starting cell population submitted for differentiation?
  A: To ensure optimal differentiation outcomes, we recommend starting with a well-characterized population of pluripotent stem cells, such as iPSCs or ESCs. These cells should exhibit robust growth characteristics, high viability, and a stable karyotype. Additionally, we advise our clients to provide detailed information regarding the passage number, culture conditions, and any prior treatments or modifications performed on the cells, enabling us to tailor the differentiation protocol accordingly.
• Q: Can detailed information be provided on the culture media and supplements used in the differentiation process?
  A: Certainly. The culture media and supplements play a crucial role in guiding pluripotent stem cells towards motor neuron differentiation. We utilize a proprietary blend of basal media supplemented with growth factors, neurotrophic factors, and small molecules known to influence neural induction and motor neuron specification. These components are carefully selected and optimized to create an environment conducive to efficient and robust differentiation, promoting the acquisition of motor neuron identity and functionality.
• Q: What methods do you employ to assess the maturity and functionality of differentiated motor neurons?
  A: Assessing the maturity and functionality of differentiated motor neurons is essential to ensure their suitability for downstream applications. We utilize a combination of morphological, molecular, and functional assays to characterize the maturity and functionality of the motor neurons. These include morphological analysis of neurite outgrowth and branching, gene expression profiling of motor neuron-specific markers, and functional assays assessing synaptic connectivity, excitability, and neurotransmitter release.

Scientific Resources

Molecular and Cellular Neuroscience

References

1. Stifani, N. Motor neurons and the generation of spinal motor neurons diversity. Frontiers in cellular neuroscience. 2014, 8, 293.
2. Boillée, Séverine, Christine Vande Velde, and Don W. Cleveland. "ALS: a disease of motor neurons and their nonneuronal neighbors." Neuron 52.1 (2006): 39-59.
3. Xu, He Jax, et al. "Generation of functional posterior spinal motor neurons from hPSCs-derived human spinal cord neural progenitor cells." Cell Regeneration 12.1 (2023): 15.

For Research Use Only. Not For Clinical Use.