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Title: Optimization of blood handling and peripheral blood mononuclear cell cryopreservation of low cell number samples.
Author: Hope, C.
Huynh, D.
Wong, Y.
Oakey, H.
Perkins, G.
Nguyen, T.
Binkowski, S.
Bui, M.
Choo, A.
Gibson, E.
Huang, D.
Kim, K.
Ngui, K.
Rawlinson, W.
Sadlon, T.
Couper, J.
Penno, M.
Barry, S.
Institutional Author: ENDIA Study Group
Issue Date: 2021
Publication Title: International Journal of Molecular Sciences
Volume: 22
Issue: 17
Start Page: 9129
Abstract: Background: Rural/remote blood collection can cause delays in processing, reducing PBMC number, viability, cell composition and function. To mitigate these impacts, blood was stored at 4 °C prior to processing. Viable cell number, viability, immune phenotype, and Interferon-γ (IFN-γ) release were measured. Furthermore, the lowest protective volume of cryopreservation media and cell concentration was investigated. Methods: Blood from 10 individuals was stored for up to 10 days. Flow cytometry and IFN-γ ELISPOT were used to measure immune phenotype and function on thawed PBMC. Additionally, PBMC were cryopreserved in volumes ranging from 500 µL to 25 µL and concentration from 10 × 106 cells/mL to 1.67 × 106 cells/mL. Results: PBMC viability and viable cell number significantly reduced over time compared with samples processed immediately, except when stored for 24 h at RT. Monocytes and NK cells significantly reduced over time regardless of storage temperature. Samples with >24 h of RT storage had an increased proportion in Low-Density Neutrophils and T cells compared with samples stored at 4 °C. IFN-γ release was reduced after 24 h of storage, however not in samples stored at 4 °C for >24 h. The lowest protective volume identified was 150 µL with the lowest density of 6.67 × 106 cells/mL. Conclusion: A sample delay of 24 h at RT does not impact the viability and total viable cell numbers. When long-term delays exist (>4 d) total viable cell number and cell viability losses are reduced in samples stored at 4 °C. Immune phenotype and function are slightly altered after 24 h of storage, further impacts of storage are reduced in samples stored at 4 °C.
Description: Includes data from BHS
Internal ID Number: 01875
Health Subject: PBMC
Type: Journal Article
Appears in Collections:Research Output

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