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Comparative effect of human albumin formulations on functional activities of cryopreserved PBMC
Author(s) -
Moskowitz Keith A,
Hickman Scott,
Meixell Ron,
Jeffrey Lisa,
Manak Mark
Publication year - 2008
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.22.1_supplement.676.5
Subject(s) - elispot , peripheral blood mononuclear cell , cryopreservation , chemistry , human serum albumin , albumin , cytokine , immunology , andrology , medicine , biology , biochemistry , in vitro , embryo , microbiology and biotechnology
Cryopreservation (CP) of peripheral blood mononuclear cells (PBMC) to remain functional upon thawing is important for their use in immunotherapy studies. Sera are often used for CP, but demonstrate variability and are replete with growth factors that can result in high backgrounds during cytokine assays. We compared two human serum albumins (HSA) [Grifols® (G) and Bayer® (B)] in the formulation of PBMC CP media containing DMSO (CryoMedia™), and monitored post‐thaw recovery, viability, and secretion of interferon‐γ (IFN‐γ) by ELISpot in response to PHA, CMV and CEF peptide pool stimuli. PBMCs from a leukopack were washed then cryopreserved at 1.5×10 7 /ml; viability prior to CP was approx. 98%. After storage in LN2 vapor mean recoveries were 80.4 (95% CI 55.6–105.2) and 68.0 (95% CI 64.4–71.6) for G and B, respectively, (p=0.4). Post‐thaw viabilities were 92% and decreased respectively to 74 % and 79% for G and B after 48 hours. In response to PHA stimuli, mean IFN‐γ ELISpot response was 1024 (95% CI 996–1051) and 436 (95% CI 407–46) SFC/well using PBMC formulated in B and G HSA, respectively (p < 0.001). Differences were also observed in the response to CMV peptide stimuli using B [18.4 (95% CI 15.7–21.2)] and G [15.4 (95% CI 13.3–17.5)], p < 0.05. Response to CEF peptides for both formulations was similar [16.6 (95% CI 13.3–20.0), B] and [17.4 (95% CI 14.7–20.1), G], p = 0.7. Background to media containing no stimuli were <1 SFC/well for both. The data suggest differences in HSA formulations used during CP of PBMC may elicit significantly different cellular immune responses to select peptide antigens. Additional PBMC donors and studies of proliferation and apoptosis are in process to further elucidate the functional effects of specific manufactures’ HSA during PBMC CP.