B-LCL-HROC32 (Bc HROC32)

B-LCL-HROC32 (Bc HROC32) is an Epstein-Barr virus (EBV)-immortalized human B lymphoblastoid cell line established from B lymphocytes isolated from either tumor tissue or peripheral blood of an adult patient. The cells were generated by ex vivo infection with EBV-containing supernatant derived from the B95/8 marmoset cell line in the presence of cyclosporin A to suppress T- and NK-cell outgrowth. Following several weeks of culture, stable lymphoblastoid outgrowth was achieved, resulting in a continuously proliferating monoclonal or oligoclonal B-cell population suitable for long-term in vitro expansion.

Immunophenotypically, B-LCL-HROC32 (Bc HROC32) exhibits a mature and activated B-cell profile characterized by expression of CD19 and CD20, along with high levels of activation and maturation markers such as CD23 and CD80. Strong expression of MHC class I and class II molecules indicates preserved antigen-presenting capacity. Depending on the individual clone, variable expression of differentiation-associated markers such as CD27, CD38, or CD138 may be observed, reflecting different stages of B-cell maturation. The cells are negative for T-cell markers, confirming lineage specificity.

Functionally, B-LCL-HROC32 (Bc HROC32) secretes immunoglobulin of a defined isotype (e.g., IgG, IgM, or IgA), which remains stable during prolonged culture. The secreted antibodies can be collected from culture supernatants and used for downstream applications, including antigen-binding assays, tumor cell recognition studies, or identification of disease-associated antigens. As an EBV-immortalized B-cell model, B-LCL-HROC32 (Bc HROC32) provides a robust in vitro platform for investigating humoral immune responses, B-cell activation and differentiation, and antibody-mediated mechanisms in the context of tumor immunology or systemic immune responses.

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.