An alternative method of panning for rat B lymphocytes.
A new method of panning for B lymphocytes is described in which the ability of the sIg+ cells to adhere depends on the nature and concentration of nonspecific protein used rather than on the use of anti-immunoglobulin. Rat lymph node cells were suspended in 3% bovine serum albumin in Tris-buffered Hanks' and incubated in tissue culture flasks to allow adherence to the plastic. The recovered bed of adherent cells was shown by flow cytometry to be greater than 90% surface immunoglobulin positive and MHC class II positive while containing very few T cells. This adherent fraction was subsequently treated with anti-T cell antibody plus baby rabbit complement to produce a highly purified sIg+ cell population containing no detectable T cells. The sIg+ cells obtained by this panning procedure were functionally active in BCGF and BCDF assays. This method provides an easy and inexpensive alternative to conventional panning with anti-immunoglobulin and also eliminates the possibility of B cell activation by exposure to anti-immunoglobulin-coated surfaces.
D000818:Animals; D000936:Antigen-Antibody Complex / Q000494:pharmacology*; D001402:B-Lymphocytes / Q000166:cytology* / Q000276:immunology; D002469:Cell Separation / Q000379:methods*; D003470:Culture Media; D005434:Flow Cytometry; D007164:Immunosorbents / Q000494:pharmacology*; D008198:Lymph Nodes / Q000276:immunology; D010641:Phenotype; D051381:Rats; D013696:Temperature
D000936:Antigen-Antibody Complex; D003470:Culture Media; D007164:Immunosorbents