Cryopreserved adult and neonatal hepatocytes require special handling to ensure optimal viability following the freeze/thaw process. Human liver stromal cells improve metabolic and transport function as well as longevity in culture of cryopreserved Human hepatocytes beyond 40 days in culture.
Please follow these methods closely to ensure the best performance of Human hepatocytes in co-culture with Human liver stromal Cells.
Required Equipment:1. All procedures should be conducted inside a biological safety cabinet such as a NuAire Class II Type A2
2. Incubator should be controlled for humidity, temperature and CO2 concentration such as a NuAire 4950 with the following settingsa. 6% CO2b. 95% humidityc. 37°CNote: It is best to fyrite incubators regularly to ensure appropriate gas concentration is maintained.
Required Supplies:1. Fresh or cryopreserved Human hepatocytes2. Cryopreserved growth arrested Human liver stromal cells3. Cryopreserved Hepatocyte Recovery Media4. Hepatocyte Plating Media5. Hepatocyte Culture Medium6. Collagen I coated plates 7. Liver biomatrix coated plates
Storage and Stability:Store the complete medium at 4°C in the dark until ready for use.
