For Research Use Only

Rat Primary Hepatocytes 
Catalog No. RA-6224F
    
Suggested Medium: Hepatocyte Medium /w Kit (500 ml)
Catalog No. M1265 

Product Description
Rat Primary Hepatocytes are derived from the liver of laboratory Sprague–Dawley rat. Cells are freshly isolated and frozen at passage 0. Each vial contains 3 x106 cells. Cells are negative for bacteria, yeast, fungi, and mycoplasma. Rat Primary Hepatocytes are tested for expression of markers using antibodies, ZO-1 by Immunofluorescence Staining. Cells can be expanded on multiwell culture plates ready for experiments under the cell culture conditions specified by Cell Biologics.

Laboratory Applications
Rat Primary Hepatocytes can be used in standard biochemical procedures include PCR, Western blotting, immunoprecipitation, metabolism, drug-drug interaction, drug transporter activity toxicity of drug candidates or cell derivatives for desired research applications.

Authorized Uses of Cell Biologics’ Products
Rat Primary Hepatocytes from Cell Biologics are distributed for internal research purposes only. Our products are not authorized for human use, for in vitro diagnostic procedures or for therapeutic procedures. Transfer or resale of any Cell Biologics’ cells or products from the purchaser to other markets, organizations or individuals is prohibited by Cell Biologics, without the company’s written consent. Cell Biologics’ Terms and Conditions must be accepted before submitting an order.

Disclaimer
Investigators should handle the cells that they receive from Cell Biologics with caution and treat all animal cells as potential pathogens, since no test procedure can completely guarantee the absence of infectious agents.

Warranty and Liability
Cell Biologics’ guarantee applies only to your purchase of Cell Biologics’ cells with Cell Biologics’ Media and Coating Solution for appropriate cell culture and cell testing following Cell Biologics’ online protocols within 35 days from the date of product delivery.

Primary Cell Culture Protocol

All cell culture procedures must be conducted in a bio-safety cabinet.
Any and all media, supplements, and reagents must be sterilized by filtration through a 0.2 µm filter. 
Use aseptic technique to prevent microbial contamination.

Medium:
Review the information provided on the Cell Biologics website about appropriate culture media (e.g. serum and other supplements). Use pre-warmed (37°C) cell culture media (30-50 ML, Catalog No. M1265) to seed cells and when changing media.

Coating of Cell Culture Plates or Dishes:
Coat sterile culture dishes or flasks with Gelatin Coating Solution (Catalog No. 6950, Cell Biologics) for 1 min, then aspirate the excess solution.

Handling of Arriving Live Cells
When you receive the live cells in the cell culture medium
•	Centrifuge cells at 50-60g for 4 minutes.
•	Aspirate the supernatant and resuspend the cell pellet in Cell Culture Medium (Catalog No. M1265, Cell Biologics).
•	Seed cells gently in plates or dishes coated with Gelatin Coating Solution (Catalog No. 6950, Cell Biologics). Cell seeding density is as below recommended (6 million cells per vial (1 ml) are seeded into 2 wells of a 6-well plate, or 1 of 60 mm plate).
•	Place dishes or plates in a humidified, 5%-CO2 incubator at 37°C for 6-12 hours.
•	Wash cells once with PBS and add the fresh culture medium until experiments.
•	Cells should be checked under a microscope to verify appropriate cell morphology.

Cell Recovery from Cryovial
•	Quickly thaw cells in cryo-vial by incubating them in a 37°C water bath for <1 min until there is just a small bit of ice left in the vial.
•	Promptly remove the vial and wipe it down with 70% ethanol.
•	Transfer cells from the vial (1 ml) to a sterile tube and add 5-6 ml of pre-warmed Cell Biologics’ Cell Culture Medium.
•	Carefully pour cell suspension into dishes or plates as recommended seeding numbers (6 million cells per vial (1 ml) are seeded into 2 wells of a 6-well plate, or 1 of 60 mm plate).
•	Note! Do not centrifuge cells.
•	Change culture media after cells attach to the culture dishes (usually it will take 6-12 hours after seeding the cells). 

Recommended Fresh-Cell Seeding:
6 million cells per vial (1 ml) are seeded into 2 wells of a 6-well plate, or 1 of 60 mm plate.

Note: The numbers of seeding cells in each well may need to be modified according to user’s experience.

•	Please send us the cell images (>80-90% confluence) if you have any question or problem with cultured cells.