B-LCL-HROG02 Cells
General information
| Description | B-LCL-HROG02 is a human Epstein-Barr virus (EBV)-transformed B lymphoblastoid cell line (B-LCL) established from the peripheral blood of a 68-year-old Caucasian male gastric cancer patient within the HROG biobank series, the gastric cancer counterpart of the HROC colorectal cancer B-LCL collection. EBV-mediated in vitro immortalization of primary peripheral blood B lymphocytes generated a stably proliferating suspension cell line with lymphoblast-like morphology. HROG02 cells constitutively express EBV latency antigens (EBNA-1, -2, -3, LMP-1, -2) and high levels of HLA class I and II molecules, co-stimulatory molecules CD80 and CD86, adhesion molecules CD54 and CD58, and B cell markers CD19, CD20, and CD23. B-LCL-HROG02 is applied in T cell and NK cell functional assays, HLA typing, mixed lymphocyte reactions, antigen presentation studies, and CTL induction protocols. Within the HROG patient-matched biobank, it provides a defined immunological reference for gastric cancer research, enabling autologous or allogeneic studies of tumor-associated immune responses. The line supports investigations of gastric cancer-specific T cell reactivity and patient-matched immune assay systems relevant to gastric tumor immunology. |
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| Organism | Human |
| Tissue | Peripheral blood |
| Disease | EBV-transformed B lymphoblastoid cell line (B-LCL); gastric cancer-associated donor |
| Applications | Immunology research; T cell and NK cell assays; HLA typing; antigen presentation studies; mixed lymphocyte reactions; CTL assay target cells; gastric cancer immunology |
Characteristics
| Age | 68 years |
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| Gender | Male |
| Ethnicity | Caucasian |
| Morphology | Lymphoblast-like |
| Cell type | B lymphoblast |
| Growth properties | Suspension |
Regulatory Data
| Citation | B-LCL-HROG02 (Cytion catalog number 302106) |
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| Biosafety level | 2 |
| NCBI_TaxID | 9606 |
| GMO Status | GMO-S2: This B-LCL contains a stably maintained EBV episome encoding viral latent-phase genes (EBNA-1/-2/-3, LMP-1/-2). EBV is classified as a risk group 2 pathogen. Handling requires BSL-2 containment. This classification applies within Germany; regulations may differ in other jurisdictions. |
Biomolecular Data
| Viruses | Transformant: EBV |
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Handling
| Culture Medium | RPMI 1640, w: 2.0 mM stable Glutamine, w: 2.0 g/L NaHCO3 (Cytion article number 820700a) |
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| Supplements | Supplement the medium with 15–20% FBS |
| Doubling time | 24 to 48 hours |
| Subculturing | Dilute the culture to 2–5 × 10⁵ cells/ml with fresh pre-warmed complete medium every 2–3 days. Count viable cells before passaging to maintain optimal density. Do not allow cultures to exceed 2 × 10⁶ cells/ml. Centrifugation (300 × g, 5 min) may be used to exchange medium when needed. No enzymatic dissociation is required. |
| Split ratio | 1 to 4 |
| Seeding density | 2 to 5 × 10⁵ cells/ml |
| Fluid renewal | Every 2 to 3 days |
| Freeze medium | As a cryopreservation medium, we use complete growth medium + 10% DMSO for adequate post-thaw viability. |
| Thawing and Culturing Cells |
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| Incubation Atmosphere | 37°C, 5% CO2, humidified atmosphere. |
| Shipping Conditions | 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. |
| Storage Conditions | 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. |