Product Details

• Catalog Number: T0262
• Unit Size: 1×10⁶ cells / 1.0 mL
• Species: Human (Homo sapiens)
• Tissue: Brain (Cerebral Microvasculature)
• Donor Information: Not disclosed
• Cell Type: Immortalized Endothelial Cells
• Morphology & Growth Properties: Adherent, polygonal
• Biosafety Level: BSL-2
• Storage: Vapor phase of liquid nitrogen or below -130°C
• Shipping: Shipped on dry ice
• Format: Cryopreserved frozen cells

Overview

Immortalized Human Cerebral Microvascular Endothelial Cells (HCMEC-Ras) serve as an advanced in vitro model for studying the blood-brain barrier (BBB) and neurovascular research. By integrating Ras gene expression, these cells exhibit prolonged proliferation and stable endothelial characteristics, making them ideal for studies on neuroinflammation, cerebrovascular permeability, and drug transport.

Key Features and Benefits

• Long-Term Stability: Immortalized for extended culture duration without senescence.
• Reliable In Vitro Model: Provides a consistent and reproducible system for BBB research.
• Optimized for Neurovascular Studies: Supports investigations into endothelial signaling, permeability assays, and cerebrovascular interactions.
• Easy to Maintain: Adapted to specific media and extracellular matrices for enhanced viability.
• Adherent Growth Properties: Facilitates straightforward culturing and experimental manipulation.

Culture & Handling Guidelines

Recommended Culture Conditions

• Coating: Use PriCoat™ T25 Flasks (G299) or Applied Cell Extracellular Matrix (G422) for optimal adhesion.
• Growth Medium: PriGrow I (TM001) supplemented with: 
  • 10% Fetal Bovine Serum (FBS)
  • 1% Penicillin/Streptomycin Solution (G255)
• Incubation Conditions: Maintain at 37°C in a humidified atmosphere with 5% CO₂.
• Seeding Density: 10,000 – 30,000 cells/cm²

Thawing Protocol

1. Quickly thaw cells in a 37°C water bath (maximum 2 minutes). Keep the vial cap above water level to avoid contamination.
2. Disinfect the vial with 70% ethanol and transfer to a biosafety cabinet.
3. Add thawed cells to a sterile 15 mL conical tube containing 5 mL of pre-warmed complete growth medium.
4. Centrifuge at 125xg for 5-7 minutes.
5. Aspirate the supernatant, gently resuspend the cell pellet in fresh complete medium, and seed into a pre-coated T25 flask.
6. Incubate under recommended conditions and allow cells to recover before passaging.

Subculturing Guidelines

1. Aspirate 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 125xg for 5 minutes, aspirate the supernatant, and resuspend the pellet in fresh growth medium.
5. Seed cells at the appropriate density into new culture vessels.
6. Incubate under recommended conditions and monitor for confluency.

Cryopreservation Guidelines

• Use Cryopreservation Medium (TM024) or complete growth medium supplemented with 10% DMSO.
• Freeze at a controlled rate (-1°C per minute) before transferring to liquid nitrogen storage.

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Disclaimer

1. This product is intended for research use only and not for human or animal therapeutic, diagnostic, or clinical applications.
2. The end-user must validate suitability for their specific application.
3. Cells must be handled under proper biosafety protocols.
4. Sales are final and subject to the completion of a Material Transfer Agreement (MTA) when applicable.
5. Product specifications may vary based on culture conditions and lab techniques.