We get only about 40% survival of our human PBMC stored at -150°C (in 90%FCS, 10% DMSO, Mr Frosty 1°C/min cooling in -80°C freezer).
In contrast with human MSC, we have over 90% survival.
Any suggestions?
Look, there's a selection through sperm which could be used to freeze according to research which has almost the same characteristics of semen; but unfortunately I have no financial support in my country nor the means to use it and investigate. In conclusion ... if it is possible to improve the survival of sperm postcongelados especially vitrified.
Regarding PBMC optimization I would expand upon Enrico's suggestion. Compare cryopreservation in different percentages of DMSO (5-15%). Add DMSO in 2 or 3 steps. Also vary the FCS from 10 to 90% in culture medium (alpha medium, DMEM, or RPMI). Then pick the best condition. I use a control rate freezer with induced nucleation so what works for me is not necessarily applicable to your Mr. Frosty technique. You should be able to achieve 60-70 percent viable recovery without much difficulty. 90% plus will take some work.
I would recommend to try to add both an intra- and extracellular stabilizing agent, e.g. 5% DMSO with 5% HES (hydroxyethylstarch). This combination is in general used in many PBMC cryopreservation protocols.
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