Cryoplateable Primary Human Hepatocytes

Product Description

The liver is a vital organ in mammals, and hepatocytes, the major cell type in liver, play critical roles in protein synthesis, detoxification of various metabolites, transformation of carbohydrates, and the production of biochemical necessary for digestion ^[1]. Primary hepatocyte culture is an important model for in vitro studies in drug development, including hepatotoxicity, drug transport, hepatitis virus infection, hepatic drug metabolism and hepatobiliary excretion ^[2].

iXCells Biotechnologies provides high quality human primary hepatocytes isolated from whole liver of human organ donors and cryopreserved on the day of isolation with ≥5 million cells in each vial. These cryoplateable primary human hepatocytes (CPHH) reach and maintain a confluent monolayer (>85% confluency) for at least 7 days, express general hepatocyte marker albumin, and show accumulation of lipid droplets. Each lot of CPHH is QC tested & validated for key Cytochromes P450 (CYPs) enzyme activity induction. They are negative for HIV-1, HBV, HCV, mycoplasma, bacteria, yeast, and fungi. CPHH cannot be subcultured or passaged.

Figure 1. Phase contrast images of CPHH. CPHH were plated in a 6-well culture dish (2 million cells/well) and cultured for 7 days. CPHH reached and maintained monolayer with >90% confluency. Hepatocytes exhibited hexagonal shape with single or double nuclei and characteristic “chicken wire” like bile canaliculus formation between hepatocytes.

Figure 2. Immunostaining and Oil Red O staining of cultured CPHH at D5. CPHH were plated in a 12-well culture dish (750,000 cells/well) and cultured for 5 days. (A) Albumin recognized by anti-human albumin antibody (green), and cytoskeleton marker F actin revealed by Alexa Fluor 555 Phalloidin (red), with cell nuclei counterstained by DAPI (blue). (B) Lipid droplet staining using Oil red O in CPHH.

Figure 3. P450 CYP activities induced by Omeprazole or Rifampin. CPHH were plated in a 12-well culture dish (750,000 cells/well) and incubated with appropriate inducers for 48 hours. Supernatant from induced and uninduced cells were analyzed using P450-Glo™ Assay (Promega, Cat. No. V8421; V8911). The fold changes of the drug induced P450 CYP enzyme activity over the basal level were calculated.

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