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Cells 2015, 4(1), 40-55; doi:10.3390/cells4010040

Serial Measurements of Apoptotic Cell Numbers Provide Better Acceptance Criterion for PBMC Quality than a Single Measurement Prior to the T Cell Assay

1
Department of Anatomy and Cell Biology, University of Wuerzburg, Koellikerstr. 6, 97070 Wuerzburg, Germany
2
Cellular Technology Ltd., 20521 Chagrin Blvd., Shaker Hts., OH 44122, USA
*
Author to whom correspondence should be addressed.
Academic Editor: Alexander E. Kalyuzhny
Received: 29 October 2014 / Accepted: 5 December 2014 / Published: 9 January 2015
(This article belongs to the Special Issue ELISPOT Research)
View Full-Text   |   Download PDF [1194 KB, uploaded 9 January 2015]   |  

Abstract

As soon as Peripheral Blood Mononuclear Cells (PBMC) are isolated from whole blood, some cells begin dying. The rate of apoptotic cell death is increased when PBMC are shipped, cryopreserved, or stored under suboptimal conditions. Apoptotic cells secrete cytokines that suppress inflammation while promoting phagocytosis. Increased numbers of apoptotic cells in PBMC may modulate T cell functions in antigen-triggered T cell assays. We assessed the effect of apoptotic bystander cells on a T cell ELISPOT assay by selectively inducing B cell apoptosis using α-CD20 mAbs. The presence of large numbers of apoptotic B cells did not affect T cell functionality. In contrast, when PBMC were stored under unfavorable conditions, leading to damage and apoptosis in the T cells as well as bystander cells, T cell functionality was greatly impaired. We observed that measuring the number of apoptotic cells before plating the PBMC into an ELISPOT assay did not reflect the extent of PBMC injury, but measuring apoptotic cell frequencies at the end of the assay did. Our data suggest that measuring the numbers of apoptotic cells prior to and post T cell assays may provide more stringent PBMC quality acceptance criteria than measurements done only prior to the start of the assay. View Full-Text
Keywords: T cell assay; apoptosis; acceptance; viability; ELISPOT T cell assay; apoptosis; acceptance; viability; ELISPOT
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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).

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MDPI and ACS Style

Wunsch, M.; Caspell, R.; Kuerten, S.; Lehmann, P.V.; Sundararaman, S. Serial Measurements of Apoptotic Cell Numbers Provide Better Acceptance Criterion for PBMC Quality than a Single Measurement Prior to the T Cell Assay. Cells 2015, 4, 40-55.

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