Related Experiment Videos
Maximizing the retention of antigen specific lymphocyte function after cryopreservation
Mary L Disis1, Corazon dela Rosa, Vivian Goodell
1Tumor Vaccine Group, Center for Translational Medicine in Women's Health, 815 Mercer Street, 2nd Floor, Box 358050, University of Washington, Seattle, WA 98195-8050, USA. ndisis@u.washington.edu
Journal of Immunological Methods
|December 13, 2005
Summary
Optimizing cryopreservation of peripheral blood mononuclear cells (PBMC) is crucial for cancer immunotherapy research. Using specific media additives and washing temperatures significantly improves PBMC viability and function after thawing.
Area of Science:
- Immunology
- Cell Biology
- Cryobiology
Background:
- Cryopreservation of peripheral blood mononuclear cells (PBMC) is essential for analyzing cancer immunotherapy.
- Maintaining lymphocyte viability and function post-thaw is critical for reliable study outcomes.
Purpose of the Study:
- To develop an optimized protocol for cryopreserving PBMC to maximize viability and function.
- To identify key factors influencing PBMC survival and function after freezing and thawing.
Main Methods:
- Evaluated various cryopreservation strategies, including different media additives and washing temperatures.
- Assessed cell viability and functional T cell responses (tetanus toxoid stimulation index) post-thaw.
- Compared outcomes using human AB serum, dextran, fetal bovine serum, and human serum albumin.
Main Results:
- Shipping method, wash/centrifugation speed, and cell number did not significantly impact viability.
- Washing media temperature critically affected viability: 4°C (69.7%), 25°C (92.55%), 37°C (95.11%).
- Human serum albumin outperformed other additives in preserving T cell function compared to dextran or fetal bovine serum; human AB serum significantly reduced viability.
Conclusions:
- Optimized cryopreservation protocols can preserve PBMC viability and function for immunotherapy studies.
- Washing media temperature and specific additives like human serum albumin are key to successful PBMC cryopreservation.
- The findings provide a robust method for cryopreserving PBMC, enhancing the reliability of cancer immunotherapy research.