Immortalized Human Cerebral Microvascular Endothelial Cells

Product Details

Catalog Number: T0259
Unit Size: 1 x 10⁶ cells / 1.0 mL
Species: Human (Homo sapiens)
Tissue of Origin: Brain
Cell Type: Immortalized Cerebral Microvascular Endothelial Cells
Growth Properties: Adherent, polygonal morphology
Immortalization Method: SV40 large T antigen transduction
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 – SV40 provide a reliable and reproducible platform for in vitro studies of brain endothelial functions. These cells have been genetically modified to express the SV40 large T antigen, allowing extended culture periods while maintaining endothelial characteristics. They serve as a valuable tool for research on neurodegenerative diseases, CNS drug permeability, and blood-brain barrier modeling.

Key Features and Benefits

Long-Term Stability: Immortalized for extended culture duration without senescence.
Consistent Reproducibility: Ensures reliable experimental outcomes.
Ideal for Neurovascular Research: Suitable for blood-brain barrier, neuroinflammation, and CNS drug transport studies.
Optimized Growth Conditions: Adapted to specific media and extracellular matrices for enhanced viability.
Adherent Growth Properties: Facilitates easy handling and maintenance in culture.

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 (G255)
Incubation Conditions: Maintain at 37°C in a humidified atmosphere with 5% CO₂.
Seeding Density: 25,000 – 30,000 cells/cm²
Doubling Time: 18 – 24 hours

Thawing Protocol

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

Subculturing Guidelines

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

Cryopreservation Guidelines

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

Disclaimer

This product is intended for research use only and is not for diagnostic or therapeutic applications.
Product performance is dependent on the end-user’s culture conditions and experimental setup.
Proper handling and laboratory safety procedures should be followed at all times.
Cells should be handled by trained personnel under aseptic conditions.
The company makes no warranties regarding the suitability of this product for specific applications beyond research use.

Reference

Yang, Sung-Hyun, et al. “Synthesis and biological evaluation of S-lipidated lipopeptides of a connexin 43 channel inhibitory peptide.” RSC Medicinal Chemistry 11.9 (2020): 1041-1047.