H4-II-E-C3

The H4-II-E-C3 cell line is a well-established rat hepatoma cell line derived from liver carcinoma. This line is widely used in research focused on liver-specific functions and the metabolic processes that occur within hepatocytes. One notable aspect of H4-II-E-C3 cells is their high expression of the high-affinity glutamate transporter GLT-1A. This feature distinguishes them from normal rat hepatocytes, which do not exhibit significant levels of this transporter. The expression of GLT-1A in H4-II-E-C3 cells allows for the efficient study of glutamate metabolism and transport, providing insights into liver cell transformation and tumor cell metabolism.

Additionally, H4-II-E-C3 cells are used to study aldehyde metabolism, particularly the activity of mitochondrial class 2 aldehyde dehydrogenase (ALDH2). This enzyme is critical for metabolizing acetaldehyde produced during ethanol oxidation, making this cell line suitable for research on alcohol metabolism and its cellular effects. The use of an "acetaldehyde clamp" technique in these cells has allowed precise measurement of ALDH2 activity without the need for mitochondrial isolation, facilitating the study of enzyme kinetics and the impact of various inhibitors on aldehyde detoxification.

37°C, 5% CO₂, humidified atmosphere.

1. Confirm that the vial remains deeply frozen upon delivery, as cells are shipped on dry ice to maintain optimal temperatures during transit.

2. Upon receipt, either store the cryovial immediately at temperatures below -150°C to ensure the preservation of cellular integrity, or proceed to step 3 if immediate culturing is required.

3. For immediate culturing, swiftly thaw the vial by immersing it in a 37°C water bath with clean water and an antimicrobial agent, agitating gently for 40-60 seconds until a small ice clump remains.

4. Perform all subsequent steps under sterile conditions in a flow hood, disinfecting the cryovial with 70% ethanol before opening.

5. Carefully open the disinfected vial and transfer the cell suspension into a 15 ml centrifuge tube containing 8 ml of room-temperature culture medium, mixing gently.

6. Centrifuge the mixture at 300 x g for 3 minutes to separate the cells and carefully discard the supernatant containing residual freezing medium.

7. Gently resuspend the cell pellet in 10 ml of fresh culture medium. For adherent cells, divide the suspension between two T25 culture flasks; for suspension cultures, transfer all the medium into one T25 flask to promote effective cell interaction and growth.

8. Adhere to established subculture protocols for continued growth and maintenance of the cell line, ensuring reliable experimental outcomes.

Mycoplasma contamination is excluded using both PCR-based assays and luminescence-based mycoplasma detection methods.

To ensure there is no bacterial, fungal, or yeast contamination, cell cultures are subjected to daily visual inspections.

For long-term preservation, place vials in vapor-phase liquid nitrogen at about −150 to −196 °C. Storage at −80 °C is acceptable only as a short interim step before transfer to liquid nitrogen.

Cryopreserved cell lines are shipped on dry ice in validated, insulated packaging with sufficient refrigerant to maintain approximately −78 °C throughout transit. On receipt, inspect the container immediately and transfer vials without delay to appropriate storage.

At Cytion, we stand behind the quality and viability of our products. We guarantee that our cells will be viable at the start of culture for 60 days after shipment from our facilities. We assure that all materials conform to the specifications described in the Certificate of Analysis and/or Product Sheet.

For full details, please refer to our Warranty Statement.