. Author manuscript; available in PMC: 2019 May 15.

Published in final edited form as: Anal Biochem. 2018 Mar 2;549:12–20. doi: 10.1016/j.ab.2018.03.003

Table 1.

Comparison Table (n=5)

	Approach 1	Approach 2			Approach 3
	(ACD/Ficoll/ PBS)	(EDTA/Ficoll/ PBS)	(EDTA/Ficoll/ DPBS)	(EDTA/Ficoll/ HBSS)	(EDTA/Lymphoprep™/ PBS)
Sample size	5	5	5	5	5
Yield (10⁶ cells/ml whole blood)	3.4± 0.2	4.1± 0.4	3.0± 0.4	2.9± 0.2	4.6± 0.3
Viability	ND	90%	ND	ND	95%
Intact cell respiration (pmol/sec/milli on cells)	3.3± 0.2	6.6± 0.5	4.4± 0.6	4.7± 0.5	7.7 ± 0.6
OXPHOS (pmol/sec/milli on cells)	8.6± 0.8	19.7± 0.9	8.4± 0.6	21.7± 1.1	23.7± 1.6

ACD: acid-citrate-dextrose; EDTA: ethylenediaminetetraacetic acid; PBS: phosphate buffered saline; DPBS: Dulbecco’s phosphate buffered saline; HBSS: Hank’s balanced salt solution; ND: Not done; OXPHOS: oxidative phosphorylation.

Whole blood has collected using vacutainers containing EDTA or ACD from single research participant. Lymphocytes were harvested with 3 different washing buffers (DPBS, PBS, and HBSS) and 2 density-gradient mediums (Ficoll-Paque and Lymphoprep™). The number of lymphocytes was counted using a hemocytometer. Viability was measured using trypan blue. Intact cell respiration and OXPHOS were measured using the Oxygraph-2K. Intact cell respiration was measured without any substrates addition. OXPHOS was determined in permeabilized cells with additions of malate (2mM), pyruvate (2.5mM), adenosine diphosphate (2.5mM), glutamate (10mM), and succinate (10mM). Data are presented as mean ± standard error (n=5).