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Immortalized Human Hepatocytes – HPV E6/E7, hTERT & MycT58A

5,800.00 USD

1×106 cells / 1.0 ml

SKU: T0063 Category: Tags: ,

Description

 

Description Hepatocytes are the chief functional cells of the liver and perform an astonishing number of metabolic, endocrine, and secretory functions. They are cuboidal epithelial cells, and 70–85% of the liver volume is occupied by hepatocytes.
Human hepatocytes are routinely used by the pharmaceutical industry for study of hepatotoxicity, drug clearance, and drug-drug interactions. However, their use is largely impeded by inadequate supply, high cost, high variability, and limited in vitro proliferation capacity. Alternative cell sources include hepatic cell lines and stem-cell derived hepatocytes but most of them do not exhibit sufficient in vivo-like functionality to be of pharmaco-toxicological relevance. In this scenario, immortalized human hepatocytes serve as the optimal in vitro model of the human liver as they are more easily maintained, propagated in culture, and have less intrinsic variation. They provide a valuable tool for investigating metabolic pathways and hepatocyte proliferation-related events.
SKU T0063
Species Human (H. sapiens)
Tissue/Organ/Organ System Liver
Growth Properties Adherent
Cell Morphology Epithelial
Seeding Density 5 – 15,000 cells/cm2; Recommended split ratio of 1:2 to 1:5*
* Do not exceed a split ratio of 1:5
Population Doubling Time 30 – 35 hours
Immortalization Method Serial passaging and transduction with recombinant lentiviruses carrying HPV E6/E7 gene
Applications For Research Use Only
Unit quantity 1×106 cells / 1.0 ml
Cell Type Immortalized Cells
Propagation Requirements Grow cells in culture vessels coated with Applied Cell Extracellular Matrix (G422). Coat plates at 37.0°C overnight and wash
with sterile PBS prior to use.

The base media for this cell line is Prigrow IX medium available at?abm?Cat. No.?TM019. To make the complete growth medium, add the following components to the base medium: fetal bovine serum (TM999)* to a final concentration of 10%, and Penicillin/Streptomycin Solution (G255) to a final concentration of 1%. Carbon dioxide (CO2): 5%, Temperature: 37.0°C.

* abm does not recommend to use heat-inactivated FBS for cell culture unless specified otherwise.

Subculture Protocol
  1. Subculture when the cells reach 80-85% confluency.
  2. Warm complete media, Trypsin-EDTA (TM050), and Dulbecco’s Phosphate Buffered Saline (Sigma) to room temperature.
  3. Wash the cells once with PBS, then add 1:10 diluted (in PBS) Trypsin-EDTA (TM050)?solution to the culture vessel. Gently rock the flask to ensure complete coverage of cells. Incubate in a 37.0°C incubator for 5 to 10 minutes.
  4. At the end of incubation, gently tap the side of the vessel to dislodge cells from the surface. If necessary, a cell scraper can be used at this point to detach cells.
  5. Add an equal volume of complete media to the vessel, then transfer suspended cells to a centrifuge tube. Rinse culture vessel with 1-2 mL complete media to collect residual cells.
  6. Centrifuge the cells at 200x g for 2-3 minutes. Remove supernatant and resuspend cells in fresh complete media.
  7. Seed cells in Collagen I, rat tail-coated culture vessel.
Preservation Protocol
  1. Freeze Medium: Complete growth medium with 20% FBS and 10% DMSO.
  2. Storage Temperature: Liquid nitrogen vapour phase.

Supporting Protocol

Other