BC-3

The BC-3 cell line is a human primary effusion lymphoma (PEL) model derived from the pleural effusion of an HIV-negative patient. It represents a rare subtype of B-cell non-Hodgkin lymphoma that typically presents as lymphomatous effusions in body cavities such as the pleura, pericardium, or peritoneum, often in the absence of detectable solid tumor masses. BC-3 is distinguished by its exclusive infection with Kaposi’s sarcoma-associated herpesvirus (KSHV or HHV-8) and complete absence of Epstein-Barr virus (EBV), a feature that sets it apart from most other PEL cell lines, which are typically dually infected.

Immunophenotypically, BC-3 cells express leukocyte common antigen (CD45) but lack classical B-cell (e.g., CD19, CD20, CD21, CD22) and T-cell (e.g., CD2, CD3, CD4, CD5, CD8) lineage markers, consistent with the undifferentiated immunophenotype of PEL. However, they do show variable expression of activation markers such as CD30, CD38, CD54, and HLA-DR. Genotypically, BC-3 exhibits clonal immunoglobulin gene rearrangements indicative of B-cell origin, but lacks c-myc oncogene rearrangements. Importantly, Southern blot and PCR analyses confirmed the presence of an intact ~170 kb KSHV genome, and electron microscopy demonstrated herpesvirus-like capsids, validating the line as a productive source of infectious KSHV particles.

BC-3 is included in the LL-100 panel of leukemia and lymphoma cell lines and clusters distinctly with other PEL models in transcriptomic analyses. It exhibits expression of PEL-characteristic genes such as PRDM1/BLIMP1, IL-10, SLAMF7, and members of the S100A family. These markers reflect a post-germinal center B-cell phenotype and underscore BC-3’s utility as a biologically relevant model for studying KSHV pathogenesis, PEL biology, and therapeutic strategies targeting KSHV-associated malignancies.

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.