A new procedure to analyze polymorphonuclear myeloid derived suppressor cells in cryopreserved samples cells by flow cytometry

PloS One
Alessandra SacchiChiara Agrati

Abstract

Myeloid derived suppressor cells (MDSC) is a heterogeneous subset of immature and mature cells of the myeloid lineage, undergoing expansion during pathologic conditions, and able to perform strong immune suppressive functions. It has been shown that cryopreservation selectively impacts the polimorphonuclear (PMN) MDSC viability and recovery, and alters the correct analysis of MDSC subsets. In laboratory practice, cryopreservation is often inevitable, in particular in multicenter studies where samples have to be shipped to a centralized laboratory. Aim of the present work was to set out a new protocol to evaluate the frequency of PMN-MDSC in thawed cells by flow-cytometry. PBMC were isolated from HIV+ patients and healthy donors, and were cryopreserved for at least ten days. After thawing, two different protocols were used: 1. standard protocol (SP) consisting of staining with the antibodies mix and then fixing with formalin 1%; 2. thawed protocol (TP) in which fixation foregoes the staining with the antibodies mix. Results showed that PMN-MDSC frequency in ex vivo PBMC evaluated by means TP was comparable to that analysed by SP, indicating that the protocol did not alter PMN-MDSC quantification in ex vivo cells. We then demonst...Continue Reading

References

Sep 6, 2003·Journal of Immunological Methods·A CostantiniM Montroni
May 4, 2007·The Journal of Clinical Investigation·Antonio Sica, Vincenzo Bronte
Feb 7, 2009·Nature Reviews. Immunology·Dmitry I Gabrilovich, Srinivas Nagaraj
Sep 29, 2011·Hepatology : Official Journal of the American Association for the Study of Liver Diseases·Robert S TackeYoung S Hahn
Apr 24, 2012·Journal of Immunological Methods·Athanasios KotsakisTheresa L Whiteside
Aug 21, 2013·Immunological Reviews·Celeste GohYoung S Hahn
Nov 27, 2014·Cytometry. Part B, Clinical Cytometry·Vera DamuzzoSusanna Mandruzzato
Dec 30, 2014·Frontiers in Oncology·Kim De VeirmanJo A Van Ginderachter
Jan 7, 2016·Journal of Translational Medicine·Eva GrütznerRika Draenert
Jun 1, 2016·Human Immunology·Peter BorosMargit Zeher
Jul 7, 2016·Nature Communications·Vincenzo BronteDmitry I Gabrilovich
Jan 6, 2017·Cancer Immunology Research·Dmitry I Gabrilovich
Jan 7, 2017·Journal of Acquired Immune Deficiency Syndromes : JAIDS·Nicola TuminoAlessandra Sacchi
Feb 10, 2017·Journal of Leukocyte Biology·Richard P TobinMartin D McCarter

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Citations

Dec 19, 2018·Pulmonary Circulation·Andrew J BryantRizwan Hamid
Mar 5, 2021·Science China. Life Sciences·Melissa S F NgLai Guan Ng
Jul 16, 2021·The Journal of Immunology : Official Journal of the American Association of Immunologists·Peter M CarlsonPaul M Sondel

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Methods Mentioned

BETA
flow cytometry
density gradient centrifugation

Software Mentioned

GraphPad Prism
FACS DIVA

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