Quantitative comparison of human intestinal mononuclear leukocyte isolation techniques for flow cytometric analyses
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Quantitative comparison of human intestinal mononuclear leukocyte isolation techniques for flow cytometric analyses. / Schreurs, R R C E; Drewniak, A; Bakx, R; Corpeleijn, W E; Geijtenbeek, T H B; van Goudoever, J B; Bunders, M J.
In: J IMMUNOL METHODS, Vol. 445, 06.2017, p. 45-52.Research output: SCORING: Contribution to journal › SCORING: Journal article › Research › peer-review
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TY - JOUR
T1 - Quantitative comparison of human intestinal mononuclear leukocyte isolation techniques for flow cytometric analyses
AU - Schreurs, R R C E
AU - Drewniak, A
AU - Bakx, R
AU - Corpeleijn, W E
AU - Geijtenbeek, T H B
AU - van Goudoever, J B
AU - Bunders, M J
N1 - Copyright © 2017 The Authors. Published by Elsevier B.V. All rights reserved.
PY - 2017/6
Y1 - 2017/6
N2 - Studies on immune cells derived from the human intestine are needed to understand the pathogenesis of gastrointestinal diseases and to develop novel treatment strategies. Isolation techniques to extract these immune cells from intestinal tissue are largely based on murine studies and comparative data on isolation from human intestine is scarce. In this study we evaluated cell yield, viability, and surface-molecule expression on mononuclear leukocytes, comparing three techniques to obtain a single immune cell suspension from human intestine; low concentrations of either the enzymes Collagenase D or Liberase TL, and enzyme-free mechanical dissociation with the Medimachine. Both enzymatic isolation techniques provided a higher cell yield than mechanical dissociation. Expression of surface molecules remained intact after Collagenase D treatment, while Liberase TL digestion resulted in a strong decrease in the expression of the CD4 receptor. Taken together, Collagenase D digestion provides the highest yield of mononuclear cells while keeping surface molecule expression intact.
AB - Studies on immune cells derived from the human intestine are needed to understand the pathogenesis of gastrointestinal diseases and to develop novel treatment strategies. Isolation techniques to extract these immune cells from intestinal tissue are largely based on murine studies and comparative data on isolation from human intestine is scarce. In this study we evaluated cell yield, viability, and surface-molecule expression on mononuclear leukocytes, comparing three techniques to obtain a single immune cell suspension from human intestine; low concentrations of either the enzymes Collagenase D or Liberase TL, and enzyme-free mechanical dissociation with the Medimachine. Both enzymatic isolation techniques provided a higher cell yield than mechanical dissociation. Expression of surface molecules remained intact after Collagenase D treatment, while Liberase TL digestion resulted in a strong decrease in the expression of the CD4 receptor. Taken together, Collagenase D digestion provides the highest yield of mononuclear cells while keeping surface molecule expression intact.
KW - Collagenases/metabolism
KW - Flow Cytometry
KW - Humans
KW - Intestinal Mucosa/metabolism
KW - Intestines/cytology
KW - Leukocytes, Mononuclear/cytology
KW - Thermolysin/metabolism
U2 - 10.1016/j.jim.2017.03.006
DO - 10.1016/j.jim.2017.03.006
M3 - SCORING: Journal article
C2 - 28274838
VL - 445
SP - 45
EP - 52
JO - J IMMUNOL METHODS
JF - J IMMUNOL METHODS
SN - 0022-1759
ER -
