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INTENDED USE
This product is for research use only (RUO). Not for use in diagnostic or therapeutic procedures.

IMPORTANT USER NOTE
These cells exhibit biological characteristics associated with the obese donor phenotype, which may impact their growth, differentiation, and metabolic behavior compared to cells from lean donors. It is the responsibility of the user to determine the suitability of this product for their specific application. Proper training in the handling of human-derived materials is required.

PRODUCT DESCRIPTION

These human spinal motor neurons (MNs) are derived from induced Pluripotent Stem Cells (iPSCs) reprogrammed from a donor with a confirmed diagnosis of sporadic Amyotrophic Lateral Sclerosis (sALS). The differentiation process utilizes a directed, floor-plate-based protocol to generate a highly pure population of cells expressing key motor neuron markers.

This product provides a physiologically relevant in vitro model sALS, which accounts for ~90% of all ALS cases. These neurons recapitulate key disease phenotypes, including increased susceptibility to cellular stress, altered mitochondrial function, and potential TDP-43 protein pathology. They are an essential tool for investigating the mechanisms of motor neuron degeneration, screening neuroprotective compounds, and developing personalized medicine approaches for ALS.

KEY FEATURES & BENEFITS

• Authentic Disease Model: Captures the complex genetic and pathological background of sporadic ALS in a human-relevant system.

• High Purity: Differentiated population is highly enriched for spinal motor neurons, expressing characteristic markers.

• Key Disease Phenotypes: Exhibits hallmarks of ALS pathology, providing a robust platform for mechanistic and therapeutic studies.

• Ready-to-Use: Cryopreserved at a post-mitotic stage, ready for functional assays upon recovery and maturation.

• Comprehensive Characterization: Each lot is validated for motor neuron identity, purity, and viability.

QUALITY CONTROL SPECIFICATIONS

Each lot is tested to meet the following release criteria:

Viability (post-thaw): ≥ 70% (Test Method: Live/Dead Staining or Trypan Blue)
Cell Yield per Vial: ≥ 2 million or 5 million viable cells (Test Method: Cell Count)
Purity (MN Marker Expression): % HB9+/Chat+ or ISL1+/Tuj1+ (Test Method: Immunocytochemistry (ICC))
Neuronal Marker Expression: Positive: β-III-Tubulin (Tuj1), MAP2; Motor Neuron Markers: ISL1, HB9 (MNX1), Chat (functional) (Test Method: ICC/Flow Cytometry)
Pluripotency Marker Absence: Negative: Oct3/4, Nanog (Test Method: ICC)
Microbiological Sterility: No growth (Test Method: USP <71>)
Mycoplasma: Negative (Test Method: PCR Method)

APPLICATIONS

• ALS Disease Modeling: Study mechanisms of motor neuron vulnerability, axonal degeneration, and glial cell interactions in co-culture.

• Neurotoxicity & Drug Screening: Identify compounds that protect against stress-induced death or modulate excitability.

• Electrophysiology: Characterize electrical properties and synaptic activity using patch-clamp or multi-electrode array (MEA) systems.

• Biomarker Discovery: Investigate protein aggregation (TDP-43), RNA metabolism, and secretory profiles.

• Gene Expression Analysis: Profile transcriptomic changes in a patient-specific background.

CELL CULTURE INFORMATION

Materials Supplied

• Product: iMN-ALSRG-S-55001 (Cryopreserved Vial)

• Components: One vial containing cryopreserved human iPSC-derived motor neurons in suspension.

Recommended Media & Reagents

• Complete Motor Neuron Maintenance Medium (Cat. No. M-4530)

• Plating Medium: Complete Maintenance Medium for the first 24-48 hours post-thaw to enhance cell survival and attachment.

• Coating Substrate: Essential. iPSC-coated solutions are required for optimal attachment and neurite outgrowth.

Thawing and Plating Procedure

Critical: Pre-coat culture vessels overnight at 4°C or 1-2 hours at 37°C, followed by rinses with PBS before plating.

1. Quick Thaw: Remove vial from liquid nitrogen and thaw rapidly in a 37°C water bath (~1-2 minutes). Do not vortex.

2. Decontaminate: Wipe the vial with 70% ethanol and transfer it to the biosafety cabinet.

3. Gentle Transfer: Using a serological pipette, gently transfer the cell suspension to a 15 mL conical tube.

4. Slow Dilution: Slowly add 5-9 mL of pre-warmed Plating Medium dropwise over 1-2 minutes to dilute the cryoprotectant gently.

5. Centrifuge: Spin at 200 × g for 5 minutes.

6. Resuspend and Plate: Aspirate supernatant. Gently resuspend the cell pellet in fresh, pre-warmed Plating Medium.

   • Recommended Seeding Density: 50,000 - 100,000 cells/cm² for high-density networks. Adjust based on application (e.g., higher for electrophysiology, lower for single-cell imaging).

7. Incubate: Plate cells immediately onto the pre-coated surface—place in a humidified 37°C incubator with 5% CO₂.

8. Medium Change: After 24-48 hours, carefully replace 50% of the medium with fresh Complete Maintenance Medium. Thereafter, perform a 50% medium change every 2-3 days.

Maintenance & Maturation

• Maturation: Cells are cryopreserved at an early post-mitotic stage. Allow 7-14 days in culture for full maturation, including robust neurite outgrowth and expression of synaptic markers.

• Handling: These are delicate, post-mitotic cells. Avoid harsh pipetting and frequent, full medium changes to prevent shear stress.

• Morphology: Expect to see phase-bright cell bodies with extensive, branching neurite networks.

Notice on Co-cultures
For co-culture with human iPSC-derived astrocytes or microglia (available separately), please don't hesitate to contact technical support for optimized protocols to ensure the proper cell ratio and medium formulation.

TYPICAL DONOR INFORMATION

• Diagnosis: Sporadic Amyotrophic Lateral Sclerosis (sALS)

• Genetics: No known mutations in SOD1, C9orf72, TARDBP, or FUS (specific details per lot)

• Age at Collection: 18 - 65 years

• Sex: Male and Female donors available

• Source Cell Type: Dermal Fibroblasts or Peripheral Blood Mononuclear Cells (PBMCs)

ORDERING INFORMATION

Required Reagents:

Essential Controls:

KEYWORDS

iPSC-derived human motor neurons for ALS research,
Patient-derived iPSC models of sporadic ALS,
Motor neuron culture for neurodegenerative drug testing,
Human stem cell-based motor neuron differentiation,
ALS sporadic iPSC-derived motor neurons,