Plating efficiency

To get the plating efficiency, murine fibroblasts (L929) are plated into a Petri dish with 20 % serum and DMEM as basal medium in a very small density (500 cells/dish). After an incubation period of 14 days in an incubator at 37° C and 5 % CO2 fumigation, the cell colonies are counted after Giemsa coloration (total plating efficiency). The results are normalized against a further tested reference serum (relative plating efficiency).

Cloning efficiency

The cloning efficiency shows the ability of a serum lot to support the cloning and the growth of murine myeloma cell line and derived hybridoma lines. Therefore SP2/0- Ag14 cells (murine myeloma) are plated on microtiter plates (one cell per well) with 20 % serum and RPMI 1640 as basal medium in a very small density. After 7 days in an incubator at 37° C and 5 % CO2 fumigation the cell colonies are counted (total cloning efficiency). The results are normalized against a further tested reference serum (relative cloning efficiency).

Growth test

In this assay the ability of a serum lot to support the proliferation of murine fibroblasts (L929) and murine myeloma cells (SP2/0-Ag14) is searched. The cells are plated at a relative low density of 1.000 cells/ml for SP2 and 10.000 cells/ml for L929. After an incubation at 37° C and 5 % CO2 fumigation cells are counted in a metering chamber on the second, fifth and seventh day. A control culture is carried along the test.