Please use this identifier to cite or link to this item:
http://hdl.handle.net/11054/1949
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 |
URI: | http://hdl.handle.net/11054/1949 |
DOI: | https://doi.org/10.3390/ijms22179129 |
Internal ID Number: | 01875 |
Health Subject: | PBMC BLOOD HANDLING CELL CONCENTRATION CRYOPRESERVATION DELAY IN PROCESSING |
Type: | Journal Article Article |
Appears in Collections: | Research Output |
Files in This Item:
There are no files associated with this item.
Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.