Question: How can I ship cells?
Answer: We recommend shipping cryopreserved cells on dry ice with enough dry ice to prevent cells from thawing.
After shipping and thawing the cells, we expect some decrease in viability and total cell recovery. The degree will primarily depend on the cell integrity and cryopreservation conditions.
Cryopreservation
Pre-cool a cell freezing container by placing it on ice before starting cryopreservation.
a. Place PBMCs on ice.
b. Gently mix the cells.
c. Determine cell viability and total cell number.
d. Centrifuge at 300 rcf for 5 min at 4°C.
e. Remove the supernatant.
f. Resuspend the cell pellet in an appropriate volume of chilled Resuspension Medium to achieve a concentration of 20,000,000 cells/ml. Maintain the cells on ice.
g. Add an equivalent volume of chilled 2X Freezing Medium to achieve a concentration of 10,000,000 cells/ml. Gently mix the cells.
h. Dispense cell suspension aliquots into the pre-cooled cryovials and place the cryovials inside a pre-cooled cell freezing container e.g., CoolCell FTS30.
i. Place the cell freezing container in a −80°C freezer for ≥4 h. After 4 h, transfer the cryovials to liquid nitrogen for long-term storage.
Thawing
This document outlines four different protocols to thaw cryopreserved cell suspensions for use with 10x Genomics Single Cell assays. One or more protocols may be applicable for a specific sample type and the most appropriate protocol should be used for thawing single cell suspensions for use with 10x Genomics Single Cell assays. This document also provides guidelines in identifying the appropriate protocol(s) by providing a comparison of the protocol features, and indicating the key differences among the four thawing protocols. The protocols described in this document are being used by 10x Genomics. Other thawing methods may also be optimized to be compatible with 10x Genomics Single Cell assays.
Other useful guidelines:
- We recommend freezing at least one million cells to ensure a sufficient number will be recovered post-thaw. It is expected that you will lose almost half from the freeze-thaw and during the wash and centrifugation steps.
-After thawing, cells should be washed and filtered (optional, recommended if debris and/or clumps are visible) and the final cell suspension counted prior to loading.
Products: Single Cell Gene Expression, Single Cell Immune Profiling