Product Details

• Catalog Number: T0251
• Unit Size: 1 x 10⁶ cells / 1.0 mL
• Species: Human (Homo sapiens)
• Tissue: Brain
• Donor Information: Female, Normal
• Cell Type: Immortalized Microglia Cells
• Growth Properties: Adherent
• Expression Markers: NGF, Iba1, CD45 and CD68
• Biosafety Level: BSL-2
• Storage: Below -130°C (vapor phase of liquid nitrogen)
• Shipping: Shipped on dry ice
• Format: Cryopreserved frozen cells

Overview

Immortalized Human Microglia – hTERT provide a stable and reproducible platform for studying microglial function in the central nervous system. Derived from primary human microglia (>99% purity), these cells maintain key microglial markers such as CD68 and NGF, allowing for extensive research into neurodegeneration, neuroinflammation, and immune-related neurological disorders.

Key Features and Benefits

• Extended Lifespan: Immortalized via hTERT to enable long-term culture without senescence.
• Stable Phenotype: Retains essential microglial markers including CD68, NGF, Iba1, and CD45.
• Reproducibility: Provides a robust model for consistent experimental outcomes.
• CNS Research-Ready: Ideal for studying neuroimmune interactions, neurodegenerative diseases, and inflammatory processes.
• Optimized Growth Conditions: Designed for compatibility with specialized microglia culture media and extracellular matrices.

Culture & Handling Guidelines

Recommended Culture Conditions

• Coating: PriCoat™ T25 Flasks (G299) or Applied Cell Extracellular Matrix (G422) for optimal adhesion.
• Growth Medium: PriGrow III (TM003) supplemented with: 
  • 10% Fetal Bovine Serum (FBS)
  • 1% Penicillin-Streptomycin Solution (G255)
• Incubation Conditions: Maintain at 37.0°C, 5% CO₂ in a humidified incubator.
• Seeding Density: 20,000 – 40,000 cells/cm²
• Doubling Time: ~24 hours

Thawing Protocol

1. Quickly thaw the vial in a 37°C water bath (max 2 min) with gentle agitation.
2. Disinfect the vial exterior with 70% ethanol and transfer to a biosafety cabinet.
3. Slowly add the cell suspension to a 15 mL conical tube containing 5 mL of pre-warmed complete growth medium.
4. Centrifuge at 125 x g for 5 minutes to pellet the cells.
5. Aspirate the supernatant, resuspend the pellet in fresh complete medium, and plate into a T25 flask.
6. Incubate under recommended conditions and allow cells to recover before passaging.

Subculturing Guidelines

• Passage cells when they reach 80% confluence.
• Detach using 0.25% Trypsin-EDTA (2-10 min at 37°C).
• Neutralize with complete medium and centrifuge at 125 x g for 5 min.
• Resuspend and seed new flasks at an appropriate split ratio (1:2 to 1:10).

Cryopreservation

• Cryopreservation Medium: PriGrow III (TM003) + 10% DMSO.
• Freezing Protocol: Slow freezing method (-1°C per minute) before transferring to liquid nitrogen storage.

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Disclaimer

1. This product is for research use only and is not intended for therapeutic, diagnostic, or clinical applications.
2. End-user must confirm suitability under their specific experimental conditions.
3. All sales are final; technical support is available for protocol optimization.
4. Availability subject to applicable institutional and regulatory requirements.