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Updated: Aug 8, 2026
Cryopreservation and Bioenergetic Evaluation of Human Peripheral Blood Mononuclear Cells
Published on: October 20, 2023
Effects of cryopreservation on immune responses: VI. An inexpensive method for freezing human peripheral blood
1Department of Medicine, Mount Sinai Hospital Medical Center/University of Health Sciences/Chicago Medical School, Illinois.
Insights
The inexpensive Mr. Frosty freezing unit effectively cryopreserves peripheral blood mononuclear cells (PBMCs), showing comparable functionality to controlled-rate freezing for immunological studies.
Area of Science:
- Immunology
- Cell Biology
- Cryopreservation Technology
Background:
- Cryopreservation is vital for preserving immune cells for research.
- Controlled-rate freezing is a standard but costly method for cell preservation.
- Inexpensive alternatives for peripheral blood mononuclear cell (PBMC) cryopreservation require validation.
Purpose of the Study:
- To evaluate the efficacy of the Mr. Frosty freezing unit for PBMC cryopreservation.
- To compare the functional recovery of PBMCs frozen using Mr. Frosty versus controlled-rate freezing.
Main Methods:
- Peripheral blood mononuclear cells (PBMCs) were cryopreserved using the Mr. Frosty unit.
- Functional assays assessed cell viability, recovery, proliferation (PHA, PWM), and cytokine secretion (IL-1, IL-2).
- Results were compared against cells frozen via controlled-rate freezing and fresh cells.
Main Results:
- PBMCs frozen with Mr. Frosty exhibited comparable viability, recovery, and proliferative responses to controlled-rate frozen cells.
- Interleukin-1 secretion was similar between Mr. Frosty and controlled-rate frozen cells.
- Immunoglobulin and IL-2 secretion were lower than controlled-rate frozen cells but comparable to fresh cells.
Conclusions:
- The Mr. Frosty unit provides a cost-effective and reliable method for cryopreserving PBMCs.
- It is suitable for immunological studies requiring functional cell recovery.
- This inexpensive method offers a practical alternative to controlled-rate freezing.
Abstract:
To determine the true usefulness of the commercially available inexpensive freezing unit (Mr Frosty) for cryopreservation of peripheral blood mononuclear cells (PBMCs), functional assays were carried out on cells that were frozen in this unit. The responses of these cells were compared with those of controlled rate frozen and fresh cells. Cell viability, recovery, proliferative responses to both phytohemagglutinin (PHA) and pokeweed mitogen (PWM), as well as interleukin (IL)-1 secreting activities of cells frozen in this container were closely comparable to controlled rate frozen cells. Although, the mean immunoglobulin (Ig) and IL-2 secreting abilities of these frozen cells were lower than the controlled rate frozen cells, these responses were still comparable to fresh cells in several of the individuals. Based on these results and considering the cost involved in controlled rate freezing method, we recommend this inexpensive unit for freezing PBMCs for subsequent immunological studies.
