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Lympho 24+ Spin Medium is a ready to use, sterile medium for preparation of Peripheral Blood Mononuclear Cell (PBMC) from 12 - 48 hour old peripheral blood, buffy coat, bone marrow, or cord blood by a simple centrifugation procedure. It reduces the RBC contamination but also reduces PBMC yield by 20%.

Most density gradient media are limited for the use of fresh whole blood. Lympho 24+ Spin Medium is recommended for the use of whole blood that is older than 8 hours. This helps to reduce the time depended contamination with granulocytes.

The enriched mononuclear cells can be used for a wide range of downstream applications such as magnetic activated cell sorting (MACS ¹) or stem cell research.

Separation scheme for PBMC enrichment with Lympho 24+ Spin Medium

At first carefully layer the sample material on top of the Lympho 24+ Spin Medium. Avoid a mixing of the two phases. Alternatively it’s possible to use pluriMate tubes. The barrier prevents a mixing of sample material and density solution. Furthermore the tube enables pouring off the enriched cells after the first centrifugation step. After the density centrifugation aspirate the upper layer (Plasma and dilution buffer). Afterwards transfer the mononuclear cell layer to a new conical tube and wash the cells.

Fig. 1: Preparation of PBMC with Lympho 24+ Spin Medium. The sample material will be overlayed on top of density gradient medium.

Fig. 2.: Layers after density gradient centrifugation. The PBMC can be found on top of the Lympho Spin Medium. Erythrocytes, Granulocytes and dead cells will pass the medium and can be found at the bottom of the tube.

¹MACS is a trademark of Miltenyi Biotec GmbH

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