Depletion of accessory cells by adherence to Sephadex G-10

Accessory cells such as macrophages and other antigen-presenting cells (APC) in suspensions of mouse spleen and lymph node are removed by means of their preferential adherence to Sephadex G-10 polymers. Unlike the use of nylon wool columns to remove B and accessory cells (UNIT 3.2), Sephadex G-10 columns are employed when the main purpose is to yield functional populations of T cells and B cells (by depleting mostly macrophages and other APC). The following technique is simple, fast, and reproducible; however, it results in some retention of B cells on the resin. The basic protocol in this unit involves assembling the column, loading and equilibrating the sterile Sephadex slurry, and passing the cells through the column followed by collection and analysis of eluate. Two support protocols describe preparation of the Sephadex G-10 slurry for use in the column by fining and sterilization, plus recycling resin from used columns.