HROHep03

HROHep03 is a human hepatocellular adenocarcinoma cell line established from the primary liver tumor of a 71-year-old Caucasian female patient within the HRO biobank series of patient-derived tumor cell lines developed by PD Dr. Michael Linnebacher since 2006. The tumor was classified as a primary adenocarcinoma at TNM stage T0NxMx, grade 3, reflecting a high-grade hepatic adenocarcinoma without confirmed distant metastasis at the time of tissue harvest. HROHep03 grows as an adherent monolayer with fibroblast-like morphology and was confirmed free of human pathogenic viruses HBV, HCV, and HIV, consistent with the stringent quality control standards of the Linnebacher biobank series.

HROHep03 is applicable in hepatocellular adenocarcinoma research, studies of high-grade liver tumor cell biology, drug sensitivity and resistance testing (sorafenib, cisplatin, 5-FU), liver tumor invasion and migration assays, and molecular pathway analysis. As part of the HRO biobank, this line provides a patient-specific biological resource that can be paired with matched immunological material from the same patient for personalized oncology research. Its fibroblast-like morphology distinguishes it phenotypically from the more common hepatocyte-like HCC lines and may reflect epithelial-to-mesenchymal features acquired during tumor progression or in vitro adaptation.

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.