LGVH - Biorepository - BLCL Collection

B95-8 Cotton-top tamarin (Saguinus oedipus) EBV B-lymphoblastoid cell line.

Reference cell-lines banked at the Viral and Human Genomics Laboratory.

Brief description

The B95-8 cell line is derived from cotton-top tamarin (Saguinus oedipus) lymphocytes, a new world (Colbian) monkey also known as bichichi or tití. The B95-8 cell line is commonly used in the field of virology and immunology as a continuous source of Epstein-Barr virus (EBV), also known as human herpesvirus 4 (HHV-4), to transform human and primate B cells into immortalized B-lymphoblastoid cell lines (BLCL’s) as a means to preserve genomic DNA traits of interest as well as to study EBV life cycle, immune responses to EBV and its associations with infectious mononucleosis, Burkitt’s lymphoma and nasopharyngeal carcinoma (Miller et al, PNAS, 1972).

The continuous and indefinite propagation of the B95-8 and BLCL’s ensures and indefinite supply of the cell line, of EBV supernatant and of BLCLs and of their genome (DNA banking applications). This cell line was distributed through the Deutsche Sammlung von Mikroorganismen und Zellkulturen (DSMZ) as catalogue number ACC 100 as well as through the American Type Culture Collection (ATCC) as catalogue number VR-1492. The European Collection of Authenticated Cell Cultures (ECACC) employed B95-8 derived EBV-transformed BLCL’s to generate in excess of 10,000 cell lines to establish the HLA typed Collection, Human Random Control DNA panels and for use in the UK human genome mapping project. EBV-transformed human cell lines remain in a state of latency producing negligible amounts of virus, the B95-8 cell line possesses the ability to re-enter the phase of active viral replication and produce large amounts of EBV particles into the tissue culture (supernatant).

The EBV particles generated by this cell line are viable and can infect human lymphocytes. EBV infection in humans can cause hematological malignancies or immune disorders. EBV is a risk group 2 pathogen and handling of B95-8 cell line, of EBV-transformed BLCL’s and tissue culture supernatant should only be carried out within a Class II biological safety cabinet (GSB) in adherence to biosafety level-3 (BSL-3) guidelines for viral production scale cultures.

Species: Cotton-top Tamarin (Saguinus oedipus) aka new world (Colbian) monkey, bichichi or tití.
Sex: Unknown
Morphology (suspension): Single or clustered round to oval lymphocytes of small and large size.
Morphology (adherent): Nearly 10% adopt a fibroblast (spindle) morphology.
Recommended growth medium: 90% RPMI-1640 w/L-glutamate + 10% to 20% fetal bovine serum (FBS).
Seed density: 0.5 million cells/ml.
Maintenance density: 0.5 to 1 million cells/ml.
Minimal density: 0.2 million cells/ml.
Cryopreservation density: 5 to 8 million cells/ml.
Cell doubling time: 41 to 72 hrs under optimal conditions.
Source tissue: Peripheral blood lymphocyte
Risk group: RG2- Epstein-Barr virus (EBV, Human herpesvirus- 4) infected cell line.
Expression profile: Expresses EBV BZLF-1 immediate early protein and capsid protein (WB and immunostaining).
Karyotype: Near diploid simian karyotype 44 (42 to 47), 5% polyploidy, exhbits chromosomal rearrangements.
Other infectious agents: Mycoplasma, HBV, HCV, HIV-1/2, HTLV-1/2 and SMRV negative (ELISA and PCR/RT-PCR).

NOTE: The cell line was originally thought to be derived from marmoset but DNA profilling has recently
confirmed a cotton-top tamarin (Saguinas oedipus) origin. Species identification also confirmed through
cytochrome C oxidase subunit 1 DNA barcoding (ANSI/ATCC ASN-0003-2015).

See associated protocol for further information

B95-8 cell line thawing, expansion, supernatant harvesting and cryopreservation in English or Español, (last updated Oct 2023).

Flask 1.0, seeded 2023/sep/20, Source cryovial B95-8-8A-P5 (2008/Sep/18)

Flask 2.0, seeded 2023/sep/20, Source cryovial B95-8-8A-P5 (2008/Sep/18) -- TERMINATED -- 2023/Sep/25

Flask 2.0, Partition 1, Day 0, x25
Flask 2.0, Partition 1, Day +1, x25
Flask 2.0, Partition 1, Day +2, x25
Flask 2.0, Partition 1, Day +5, x10
Flask 2.0, Partition 1, Day +5, x25

Flask 2.1, Partition 1, Day 5, x10
Flask 2.1, Partition 1, Day 5, x25

Flask 3.0, seeded 2023/sep/25, Source cryovial B95-8-8A-P3 (2008/Sep/08)