Product Details

• Catalog Number: T0056
• Unit Size: 1×10⁶ cells / 1.0 ml
• Species: Human (Homo sapiens)
• Tissue of Origin: Liver
• Donor Information: Adult
• Cell Type: Immortalized hepatic sinusoidal endothelial cells
• Growth Properties: Adherent, polygonal morphology
• Immortalization Method: Lentiviral transduction with SV40 Large T antigen
• Expression Markers: CK18, CK19, vWF, CD31, VE-cadherin, puromycin resistance
• Biosafety Level: BSL-2
• Storage: Below -130°C (liquid nitrogen vapor phase)
• Shipping: Shipped on dry ice
• Format: Cryopreserved frozen cells

Overview

The Immortalized Human Hepatic Sinusoidal Endothelial Cells (HSEC) provide a robust model for studying liver microvascular function, antigen presentation, and immune tolerance. These cells dynamically regulate porosity in response to zonal stimuli and play a key role in liver homeostasis and regeneration.

Key Features and Benefits

• Liver Immunology & Endothelial Research: Facilitates the study of antigen presentation and immune modulation.
• Dynamic Porosity Regulation: Responds to zonal environmental stimuli, crucial for liver function.
• Drug Discovery & Toxicology: Suitable for testing hepatotoxic compounds and vascular-targeted therapies.
• Consistent & Reproducible: Maintains a stable genetic background for reliable experimental outcomes.
• Adaptability to 3D Culture & Co-Culture: Enhances liver disease modeling and mechanistic studies.

Culture & Handling Guidelines

Recommended Culture Conditions

• Coating: Use Applied Cell Extracellular Matrix (G422). Coat plates at 37°C overnight and wash with sterile PBS prior to use.
• Growth Medium: PriGrow IX (TM019) supplemented with:
  • 10% Fetal Bovine Serum (FBS)
  • 1% Penicillin/Streptomycin Solution (G255)
• Incubation Conditions: 37°C in a humidified atmosphere with 5% CO₂
• Seeding Density: 20,000 cells/cm²
• Doubling Time: 12–22 hours

Thawing Protocol

1. Quickly thaw cells in a 37°C water bath while gently agitating the vial (maximum 2 minutes). Keep the vial cap above the water level to avoid contamination.
2. Decontaminate the vial by spraying with 70% ethanol and transfer it to a biological safety cabinet.
3. Transfer the cell suspension into a sterile 15mL conical tube containing 5mL of pre-warmed complete growth medium. Centrifuge at 125xg for 5–7 minutes.
4. Aspirate the supernatant without disturbing the cell pellet. Resuspend the pellet in fresh complete growth medium and seed into a pre-coated T25 flask.
5. Incubate under recommended conditions and allow the cells to recover before passaging.

Subculturing Guidelines

1. Aspirate the culture medium and rinse the cells with sterile PBS.
2. Add 2–3mL of pre-warmed 0.25% Trypsin-EDTA and incubate at 37°C until cells detach (~2–10 minutes).
3. Neutralize Trypsin-EDTA by adding an equal volume of complete growth medium.
4. Transfer the cell suspension to a sterile centrifuge tube and centrifuge at 125xg for 5 minutes.
5. Aspirate the supernatant and resuspend the cell pellet in fresh complete growth medium.
6. Seed at the appropriate density and incubate under recommended conditions.

Cryopreservation Guidelines

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

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Disclaimer

• This product is intended for research use only and is not approved for diagnostic, therapeutic, or clinical applications.
• Users are responsible for determining the suitability of this product for their specific application.
• Cells must be handled under Biosafety Level 2 (BSL-2) containment following institutional guidelines.
• No warranties are provided regarding performance beyond the described specifications.

References

• Jung, Hong-Ryul, et al. “Cell spheroids with enhanced aggressiveness to mimic human liver cancer in vitro and in vivo.” Scientific Reports 7.1 (2017): 10499.
• Lee, Ho-Joon, et al. “Elasticity-based development of functionally enhanced multicellular 3D liver encapsulated in hybrid hydrogel.” Acta Biomaterialia 64 (2017): 67-79.
• Buniatian, Gayane Hrachia, et al. “Antifibrotic effects of amyloid-beta and its loss in cirrhotic liver.” Cells 9.2 (2020): 452.
• Lee, Ho‐Joon, et al. “Optimization of 3D hydrogel microenvironment for enhanced hepatic functionality of primary human hepatocytes.” Biotechnology and Bioengineering 117.6 (2020): 1864-1876.