Fresh Frozen Human Peripheral Blood Mononuclear Cells for Single Cell RNA Sequencing

Demonstrating the Storage and Reconstitution of Human Peripheral Blood Mononuclear Cells

Demonstrated Protocol, Last Modified on March 31, 2021, Permalink

CG00039_Demonstrated_Protocol_FreshFrozenHumanPBMCs_RevD.pdf

20190802_CG00039_FreshFrozenHumanPBMCs_RevCtoRevD.pdf

This Demonstrated Protocol outlines (i) how to generate cryopreserved human peripheral blood mononuclear cells (PBMC) and (ii) how to thaw, wash, and count cryopreserved human PBMC in preparation for use in 10x Genomics Single Cell Protocols. While this Demonstrated Protocol is specific to PBMC, the Protocol may be used as a basis for handling other primary cells in preparation for use in 10x Genomics Single Cell Protocols.

This demonstrated protocol is optimized for counting cells in the range of 700-1200 cells/µl. If using the Single Cell 3' LT v3.1 (low throughput) application, ensure cells are counted as indicated in this protocol and then diluted to the LT specific optimal loading concentration of 100-600 cells/µl using the Cell Dilution Overview in the LT User Guide.