Single Cell Suspension From Spleen Protocol

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Single cell suspension from spleen protocol. This protocol describes how to harvest cells from a spleen sample and how to prepare a single cell suspension prior to performing cell isolation. Preparing a true single cell suspension of the primary tissue sample will optimize cell separation by avoiding additional cell loss and enabling maximum labeling of the target cells. Centrifuge the tube at 300 g at 40c for 5 min. If you are not using an isolation kit following dissociation please use pbs 1 mm edta.

Transfer the spleen to be processed into a sterile 35 mm culture dish cat 27100 27150 containing 5 ml of recommended medium for downstream isolation this will be indicated on your kit pis. How to prepare a single cell suspension from mouse spleen this video demonstrates how to harvest cells from a spleen sample and prepare a single cell suspension prior to performing cell isolation. Add 5 ml 1x pbs and mince the tissue with the syringe plunger after fine mincing place the cell strainer in the centrifuge tube use the syringe plunger to macerate the cells through the filter add 5 ml hbss. Preparing a true single cell suspension of the primary tissue sample will optimize cell separation from tissues by avoiding additional cell loss and enabling maximum labeling of the target cells.

Remove supernatant and add hbss solution to wash again.