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Quantifying the Detrimental Effects of Multiple Freeze/Thaw Cycles on Primary Human Lymphocyte Survival and Function.
Serra, Valentina; Fiorillo, Edoardo; Cucca, Francesco; Orrù, Valeria.
Affiliation
  • Serra V; Institute for Genetic and Biomedical Research, National Research Council (CNR), 08045 Lanusei, Italy.
  • Fiorillo E; Institute for Genetic and Biomedical Research, National Research Council (CNR), 08045 Lanusei, Italy.
  • Cucca F; Institute for Genetic and Biomedical Research, National Research Council (CNR), 08045 Lanusei, Italy.
  • Orrù V; Department of Biomedical Sciences, University of Sassari, 07100 Sassari, Italy.
Int J Mol Sci ; 24(1)2022 Dec 30.
Article in En | MEDLINE | ID: mdl-36614075
The use of cryopreserved peripheral blood mononuclear cells is common in biological research. It is widely accepted that primary cells are rendered unusable by several freezing cycles, although this practice might be very helpful when the biological material is valuable and its re-collection is impractical. To determine the extent to which primary cells undergoing repeated freezing cycles are comparable to one another and to fresh samples, we evaluated overall lymphocyte viability, their proliferation and cytokine production capabilities, as well as the levels of 27 cell subtypes in ten human peripheral blood mononuclear cells frozen for five years and repeatedly thawed. As expected, we observed a progressive increase in cell death percentages on three rounds of thawing, but the frequency of the main lymphocyte subsets was stable across the three thawings. Nevertheless, we observed a significant reduction of B cell frequency in frozen samples compared to fresh ones. On repeated thawings and subsequent conventional stimulation, lymphocyte proliferation significantly decreased, and IL-10, IL-6, GM-CSF, IFN-gamma, and IL-8 showed a trend to lower values.
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Full text: 1 Collection: 01-internacional Database: MEDLINE Main subject: Leukocytes, Mononuclear / Cryopreservation Limits: Humans Language: En Journal: Int J Mol Sci Year: 2022 Type: Article Affiliation country: Italy

Full text: 1 Collection: 01-internacional Database: MEDLINE Main subject: Leukocytes, Mononuclear / Cryopreservation Limits: Humans Language: En Journal: Int J Mol Sci Year: 2022 Type: Article Affiliation country: Italy