Summary
Methods are described for the efficient transformation of fresh or cryopreserved human B lymphocytes to produce continuous, lymphoblastic cell lines. Lymphocytes are separated from whole blood by centrifugation through Ficoll. They are transformed by exposure to Epstein-Barr Virus (EBV) obtained as a supernatant from ATCC. CRL 1612 (B95-8) cells. Virus production is verified in advance by immunoperoxidase staining after application of a monoclonal antibody to EBV capsid antigen [ATCC.HB 168 (72A1)]. The addition of irradiated feeder cells [ATCC.CCL 17 (MRC-5)] is important to enhance efficiency in lymphoblast culture initiation.
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Caputo, J.L., Thompson, A., McClintock, P. et al. An effective method for establishing human B lymphoblastic cell lines using epstein-barr virus. Journal of Tissue Culture Methods 13, 39–44 (1991). https://doi.org/10.1007/BF02388202
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DOI: https://doi.org/10.1007/BF02388202