Amyotrophic Lateral Sclerosis (ALS) In Vitro Modeling Service

Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disorder characterized by the selective degeneration of upper and lower motor neurons, leading to muscle denervation and respiratory failure. Unlike Alzheimer's disease (AD) or Parkinson's disease (PD), ALS pathology is driven by TDP-43 proteinopathy, C9orf72 repeat expansions, and non-cell autonomous toxicity from glial cells.

Creative Biolabs' ALS cellular models prioritize TDP-43 proteinopathy, C9orf72 repeat expansion toxicity, astrocyte-driven excitotoxicity, and neuromuscular junction (NMJ) dysfunction, to help scientists to screen compounds or ASOs.

Core Pathological Hallmarks & Cellular Targets

ALS pathology involves several key mechanisms:

• TDP-43 mislocalization: Cytoplasmic aggregation of TAR DNA-binding protein 43 disrupts RNA metabolism and axonal transport.
• C9orf72 dipeptide repeats (DPRs): Toxic poly-GA/GR proteins from GGGGCC hexanucleotide repeats induce nucleolar stress and proteasome dysfunction.
• Accumulation of misfolded SOD1: Mutations in superoxide dismutase 1 (SOD1) drive familial ALS by promoting misfolded protein aggregates that induce mitochondrial dysfunction, endoplasmic reticulum stress, and excitotoxicity via impaired glutamate reuptake in motor neurons.
• Cytoplasmic aggregation of mutant FUS: Mutations in RNA-binding protein Fused in Sarcoma (FUS) cause nuclear-to-cytoplasmic mislocalization, forming stress granule-like aggregates that sequester RNA-binding proteins and impair mRNA splicing/transport.
• Astrocyte-mediated excitotoxicity: Mutant SOD1 or C9orf72 astrocytes reduce glutamate uptake via EAAT2 downregulation, leading to motor neuron hyperexcitability.
• Axonal transport defects: Impaired mitochondrial trafficking in motor neuron axons.

Key cellular targets:

• Motor neurons: ALS primarily targets upper and lower motor neurons, which progressively degenerate and die in ALS, leading to muscle weakness and paralysis.
• Skeletal muscle cells: Each skeletal muscle cell connects with a motor neuron at a specialized synapse called the neuromuscular junction (NMJ). Loss of synapses at the NMJ is one of the first detectable pathological changes in ALS.
• Glial cells: Astrocyte and microglia dysfunction leads to reduced motor neuron survival.

Validated In Vitro Model Systems

Creative Biolabs offers all the validated in vitro models listed below, providing versatile tools for ALS research.

• iPSC-Derived Motor Neuron-Astrocyte/Microglia/Skeletal Muscle Cells Co-Cultures
  
  • Patient-specific lines: Derived from ALS patients with TARDBP (TDP-43), C9orf72, SOD1, or FUS mutations.
  • Isogenic controls: Utilizing CRISPR-corrected patient-derived iPSCs.
  • Healthy controls: Generated from somatic cells of healthy individuals.
• Primary Cultures derived from spinal cord or cortex
  
  • Primary neurons: Isolated from rodent or human spinal cord/cortical tissues, modeling intrinsic neuronal vulnerabilities, such as early axonal transport defects and hyperexcitability in SOD1 transgenic mice.
  • Primary astrocytes: Transgenic mice -derived astrocytes induce motor neuron toxicity via inflammatory mediators (e.g., TNF-α, IL-1β) in co-culture systems, replicating non-cell-autonomous degeneration.
  • Primary microglia: Microglia from C9orf72 transgenic mice show heightened pro-inflammatory responses to stimuli like LPS, driving synaptic pruning and neuronal loss.
• Engineered Cell Lines
  
  • TDP-43 overexpression cell line: HEK293 or SH-SY5Y cells with inducible TDP-43 expression recapitulate cytoplasmic mislocalization and stress granule dynamics.
  • TDP43-FUS stable cell lines: Co-expressing mutant TDP-43 and FUS, revealing synergistic effects on RNA metabolism and nucleocytoplasmic transport.
• Organotypic Spinal Cord/Brain Slice Cultures
  These ex vivo models preserve 3D tissue architecture and synaptic connectivity, enabling longitudinal studies of ALS progression.
  Applications:
  
  • Live imaging of calcium dysregulation and mitochondrial dysfunction in motor neurons within intact spinal cord slices.
  • Electrophysiological recordings to assess hyperexcitability in cortical slices from TDP-43 transgenic mice.
• NMJ-on-Chip Microfluidic Platforms
  Bioengineered systems integrate motor neurons and muscle cells in compartmentalized devices to model NMJ dysfunction.
  Key features:
  
  • Axon guidance channels separate neuronal somas from muscle compartments, enabling precise study of NMJ formation/regression.
  • Real-time monitoring of acetylcholine release and muscle contraction using optogenetic or microelectrode arrays.

Related Services

• ALS In Vitro Assay
  
  • Stress Granule Formation Assay
  • C9orf72 Formation Assay
  • C9orf72 Aggregation Assay
  • Cellular Localization Assay
  • TDP-43, FUS, SOD1 Assay
  • ALS High-Throughput Phenotypic Screening Assay
  • ALS Therapeutic Target Characterization Assay

Conclusion

Creative Biolabs offers a comprehensive suite of ALS in vitro models and assays designed to accelerate your research and drug discovery efforts. Our validated model systems, including iPSC-derived co-cultures, primary cultures, engineered cell lines, organotypic slices, and NMJ-on-chip platforms, provide valuable tools for studying ALS pathology and screening potential therapeutics.

Contact Us

At Creative Biolabs, we have a passionate team of scientists who are ready to provide you with expert technical support and consultation services. Don't hesitate to reach out! Contact us now to discuss your specific ALS research needs and discover how our innovative models and assays can advance your work.

Reference

1. Bonafede, Roberta, and Raffaella Mariotti. "ALS Pathogenesis and Therapeutic Approaches: The Role of Mesenchymal Stem Cells and Extracellular Vesicles." Front Cell Neurosci. 2017;11:80. Distributed under Open Access License CC BY 4.0 without modification.