Product Details

• Catalog Number: T0284
• Unit Size: 1 × 10⁶ cells / 1.0 ml
• Species: Human (Homo sapiens)
• Tissue: Brain
• Donor Information: 8-week-old fetal human ventral mesencephalon
• Cell Type: Immortalized Dopaminergic Neuronal Precursor Cells
• Growth Properties: Adherent with flat dendritic processes
• Expression Markers: DAT, VMAT-2, TH, α-SYN and β-III tubulin
• Biosafety Level: BSL-2
• Storage: Vapor phase of liquid nitrogen or below -130°C
• Shipping: Shipped on dry ice
• Format: Cryopreserved frozen cells

Overview

The Immortalized Human Dopaminergic Neuronal Precursor Cells (LUHMES) are derived from human mesencephalic tissue and provide a stable and reproducible platform for dopaminergic neuron research. These cells differentiate into a neuron-like phenotype and express key markers such as dopamine transporter (DAT), vesicular monoamine transporter (VMAT-2), tyrosine hydroxylase (TH), and β-III tubulin. Additionally, LUHMES exhibit electrophysiological properties, making them a robust in vitro model for neurodevelopment, neurodegenerative disorder research, and neuropharmacology studies.

Key Features and Benefits

• Reliable Differentiation Potential: Capable of acquiring a dopaminergic neuron-like phenotype.
• Electrophysiological Properties: Ideal for studying neuronal function and synaptic activity.
• Consistent and Reproducible Model: Provides uniform results for neurodevelopment and neurotoxicity studies.
• Long-Term Culture Stability: Immortalized via a tetracycline-controlled system to maintain cell integrity and function.
• Versatile Applications: Suitable for neurodegeneration, neuroinflammation, and pharmaceutical testing.

Culture & Handling Guidelines

Recommended Culture Conditions

• Coating: Use 50 μg/ml Poly-L-Ornithine (PLO) and 1 μg/ml Fibronectin in sterile water for at least 3 hours at 37°C before use.
• Growth Medium: 
  • Advanced DMEM/F12 (Gibco, Cat. #12634010)
  • 1X N2 Supplement (ThermoFisher Scientific)
  • 2 mM L-Glutamine (G275)
  • 40 ng/ml Recombinant Human FGF2 (Z101455)
  • 1% Penicillin/Streptomycin Solution (G255)
• Incubation Conditions: Maintain at 37°C in a humidified atmosphere with 5% CO₂.
• Seeding Density: 50,000 – 100,000 cells/cm²
• Doubling Time: 30 – 40 hours

Thawing Protocol

1. Thaw cells quickly in a 37°C water bath (~2 minutes). Keep the vial cap above the water level to avoid contamination.
2. Decontaminate the vial exterior using 70% ethanol and transfer to a biosafety cabinet.
3. Add thawed cells to a sterile 15mL conical tube containing 5mL of pre-warmed complete growth medium supplemented with 10% FBS.
4. Centrifuge at 125 × g for 5 minutes, aspirate supernatant, and gently resuspend the cell pellet in fresh complete medium.
5. Seed the cells into a pre-coated T25 culture flask and incubate under standard conditions.
6. Allow at least 4–5 days for cell attachment before the first media change.

Subculturing Guidelines

1. Aspirate the old medium and rinse cells with PBS.
2. Add 2-3 mL of pre-warmed 0.25% Trypsin-EDTA and incubate at 37°C until cells detach (~2-10 minutes).
3. Neutralize trypsin by adding an equal volume of complete medium.
4. Centrifuge at 125 × g for 5 minutes, aspirate supernatant, and resuspend pellet in fresh growth medium.
5. Seed cells at the appropriate density into new culture vessels and incubate under recommended conditions.

Cryopreservation Guidelines

• Cryopreservation Medium: Cryopreservation Medium (Cat. #TM024) or complete growth medium supplemented with 10% DMSO.
• Storage Conditions: Cells should be stored in the vapor phase of liquid nitrogen.

Related Products

• • Recombinant Human FGF2 – Z101455
  • Recombinant Human BDNF – Z100065
  • Recombinant Human GDNF – Z101055

• Recombinant Human MIF (E. coli) – Z100915

Disclaimer

• This product is for research use only and is not intended for human or animal therapeutic applications.
• All test parameters and cell characteristics are validated under standard culture conditions and may vary based on user protocols.
• A Material Transfer Agreement (MTA) may be required for purchase. Please contact customer support for further details.
• Cell viability is warranted for 30 days post-shipment, provided the product is stored and handled as per recommended guidelines.
• Users must ensure appropriate biosafety precautions when handling this product in laboratory settings.

Reference

• Scholz, Diana, et al. “Rapid, complete and large‐scale generation of post‐mitotic neurons from the human LUHMES cell line.” Journal of neurochemistry 119.5 (2011): 957-971.