Generation of cell lines using Epstein-Barr Virus (EBV) transformation of small volumes of cryo-preserved whole blood and the use of bench-top flow cytometry to achieve high and reproducible success

ECACC has developed a procedure for the generation of lymphoblastoid cell lines (LCLs) by the direct EBV transformation of small amounts (800 l) of cryo-preserved whole blood eliminating any requirement for prior separation of peripheral blood lymphocytes (PBLs). The ability to directly transform frozen whole blood avoids the cost of separating and storing PBLs while retaining the option to make a cell line at any time from all or a subset of the collection. Consistent transformation success rates of over 90% have been achieved using bench-top flow cytometry to track lymphoblastoid proliferation from source blood that has been cryo-preserved for more than four years.