So I had ordered the wrong type of DPBS and would like to know if it can still be used to perform PBMC isolations?
Normally we use DPBS without magnesium and calcium and make a 2% FBS solution with the DPBS to dilute the whole blood and when performing our washes. i know RPMI 1640 is also used in PBMC isolation and contains calcium and magnesium so could the same thing be applied to DPBS? After separation they are resuspended in freezing medium and then later placed in cryo-storage. So I'm concerned that the Mg and Ca will affect the resuspension. Has anyone encountered this issue?
Hello Tiffany
Difference between RPMI and DPBS is the pH regulation - RPMI designed to offer pH7.2 at 5.0% CO2 while DPBS at 0.037% atmospheric levels. Calcium is involved with cell vitality including enzyme function, attachment, movement, tonicity, proliferation, and metabolic processes. Removing mineral salts helps hold cell in stasis in an inactivated state.
For cryo-storage the presence of salts should not be detrimental and the cells can be transitioned to any appropriate media, following thawing, dependant on the subsequent assay requirement?
Best wishes
[Calcium in cell culture - https://www.sigmaaldrich.com/GB/en/technical-documents/technical-article/cell-culture-and-cell-culture-analysis/mammalian-cell-culture/calcium-in-cell-culture}
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