| abstract |
Herein is reported a method for obtaining a B-cell comprising the following steps a) obtaining B-cells from an experimental animal, b) labeling said B-cells, c) depositing said labeled B-cells as single cells, d) co-cultivating said single cell deposited B-cells with a feeder cell, e) selecting a B-cell proliferating and secreting IgG in step d) and thereby obtaining a B-cell. The labeling can be of IgG + CD 19 + -B-cells, IgG + CD38 + -B-cells, IgG + CD268 + -B-cells, IgG - CD138 + -B-cells, CD27 + CD138 + -B-cells or CD3 - CD27 + -B-cells. The method can comprise the step of incubating said B-cells at 37 °C for one hour in EL-4 B5 medium prior to the depositing step. The method can also comprise the step of centrifuging said single cell deposited B-cells prior to the co-cultivation. In the co-cultivation a feeder mix comprising interleukin-l beta, and tumor necrosis factor alpha and Staphylococcus aureus strain Cowans cells or BAFF or interleukin-2 and/or interleukin-10 and/or interleukin-6 and/or interleukin-4. |