Hep3B cells are a widely used human cell line in scientific research, serving as an important model for understanding liver cancer. These cells are derived from hepatocellular carcinoma, a common type of liver malignancy. Scientists employ Hep3B cells to investigate the disease’s complex biological mechanisms and explore potential therapeutic interventions. Their characteristics make them valuable in various biomedical studies.
Origin and Derivation
The Hep3B cell line was established in 1976 from tumor tissue obtained from an 8-year-old Black male patient diagnosed with hepatocellular carcinoma. The cells were isolated at Barbara B. Knowles’s laboratory within the Wistar Institute in Philadelphia, USA. This origin provides foundational context for the cell line.
Derived from a human liver tumor, Hep3B cells are relevant for studying the disease in a laboratory setting. This lineage ensures that the cells retain many biological features of the original cancer. Their establishment contributes to their reliability as a research model.
Unique Biological Characteristics
Hep3B cells possess distinct biological features. A notable characteristic is the integration of the Hepatitis B virus (HBV) genome into the cells’ DNA, meaning viral genetic material is incorporated into their human chromosomes.
Despite containing the integrated HBV genome, Hep3B cells do not produce the Hepatitis B surface antigen (HBsAg) or infectious HBV virions. This allows researchers to study viral DNA’s role in carcinogenesis without active viral replication or infectivity.
The cells also produce and secrete alpha-fetoprotein (AFP), a recognized biomarker for liver cancer often elevated in patients with hepatocellular carcinoma. Hep3B cells exhibit an epithelial-like morphology, meaning they grow as flattened, tightly packed cells, similar to the epithelial cells found in liver tissue.
Applications in Biomedical Research
Hep3B cells make them an important in vitro model for studying hepatocellular carcinoma (HCC). Researchers use them to investigate the molecular mechanisms of HCC development and progression, including genetic mutations, cellular processes, and signaling pathways.
They are employed for screening potential anti-cancer drugs and therapies. Scientists evaluate the efficacy and toxicity of new compounds against liver cancer cells in a controlled laboratory environment. Studies have explored the effects of various agents, including natural extracts and targeted inhibitors, on Hep3B cell growth and survival.
The integrated HBV genome also makes Hep3B cells suitable for Hepatitis B virus research. This feature helps scientists understand how viral DNA integration contributes to liver cancer development and provides insights into the long-term consequences of HBV infection.
Laboratory Culturing and Handling
Maintaining Hep3B cells in a laboratory requires specific conditions for proper growth. They are cultured in Eagle’s Minimum Essential Medium (EMEM), a nutrient-rich solution, supplemented with 10% fetal bovine serum (FBS) for essential growth factors and proteins.
Additional components like L-glutamine, non-essential amino acids, and sodium bicarbonate are included to meet the cells’ metabolic needs. Hep3B cells thrive in a humidified incubator at 37°C. A controlled atmosphere of 5% carbon dioxide (CO2) is necessary to regulate the medium’s pH and support cellular functions.
These cells exhibit adherent growth, attaching to cell culture flasks. For routine maintenance, confluent cultures are split every 3-5 days at a ratio of 1:2 to 1:6, using enzymes like trypsin-EDTA to detach them. Their doubling time is approximately 36 to 50 hours.