Density separation of spleen cells increases fusion frequency and yield of Ig-producing hybridomas.
The efficiency of hybridoma formation and growth after cell fusion can be much improved by fractionation of the mouse splenocytes. A simple procedure is described in which splenocytes with a specific gravity of more than 1.065 g/cm3 are selected by centrifugation on a Percoll gradient. The resulting cell suspension is largely depleted of macrophages and fibroblasts while the cell viability is improved. In fusion experiments performed with these cells, overgrowth of hybridomas by macrophages, fibroblasts and P-cells is avoided. The fusion efficiency and the frequency of immunoglobulin-secreting hybridomas is increased compared with fusions carried out with unfractionated spleen cells.
D000818:Animals; D000921:Antibody-Producing Cells / Q000166:cytology / Q000276:immunology*; D002459:Cell Fusion; D002469:Cell Separation / Q000379:methods*; D002470:Cell Survival; D002499:Centrifugation, Density Gradient / Q000379:methods; D006801:Humans; D006825:Hybridomas / Q000166:cytology / Q000276:immunology*; D007136:Immunoglobulins / Q000096:biosynthesis; D051379:Mice; D008807:Mice, Inbred BALB C; D011205:Povidone; D012822:Silicon Dioxide; D013154:Spleen / Q000166:cytology / Q000276:immunology
D007136:Immunoglobulins; C016039:Percoll; D012822:Silicon Dioxide; D011205:Povidone