Human serum albumin and chromatin condensation rescue ex vivo expanded γδ T cells from the effects of cryopreservation
| Autor: | Donald Tope, Branella Gianna, Erich Williams, H. Trent Spencer, Christopher B. Doering, Rebecca E. Burnham |
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| Rok vydání: | 2020 |
| Předmět: |
Cell type
Cell Survival T cell T-Lymphocytes Cell Serum Albumin Human General Biochemistry Genetics and Molecular Biology Cryopreservation Article 03 medical and health sciences 0302 clinical medicine In vivo Freezing medicine Humans Viability assay 030219 obstetrics & reproductive medicine Chemistry 0402 animal and dairy science 04 agricultural and veterinary sciences General Medicine 040201 dairy & animal science Chromatin Cell biology medicine.anatomical_structure Apoptosis General Agricultural and Biological Sciences Ex vivo |
| Zdroj: | Cryobiology |
| ISSN: | 1090-2392 |
| Popis: | Clinical applications of gamma delta (γδ) T cells have advanced from initial interest in expanding γδ T cells in vivo to the development of a manufacturing process for the ex vivo expansion. To develop an "off-the-shelf" allogeneic γδ T cell product, the cell manufacturing process must be optimized to include cryopreservation. It is known that cryopreservation can dramatically reduce viability of primary cells and other cell types after thawing, although the exact effects of cryopreservation on γδ T cell health and functionality have not yet been characterized. Our aim was to characterize the effects of a freeze/thaw cycle on γδ T cells and to develop an optimized protocol for cryopreservation. γδ T cells were expanded under serum-free conditions, using a good manufacturing practice (GMP) compliant protocol developed by our lab. We observed that cryopreservation reduced cell survival and increased the percentage of apoptotic cells, two measures that could not be improved through the use of 5 GMP compliant freezing media. The choice of thawing medium, specifically human albumin (HSA), improved γδ T cell viability and in addition, chromatin condensation prior to freezing increased cell viability after thawing, which could not be further improved with the use of a general caspase inhibitor. Finally, we found that cryopreserved cells had depolarized mitochondrial membranes and reduced cytotoxicity when tested against a range of leukemia cell lines. These studies provide a detailed analysis of the effects of cryopreservation on γδ T cells and provide methods for improving viability in the post-thaw period. |
| Databáze: | OpenAIRE |
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