Quantitative comparison of human intestinal mononuclear leukocyte isolation techniques for flow cytometric analyses
Open accessAbstract
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.
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Enzymatic digestion causes substantial loss of some cell-surface receptors.
“Of note, we observed substantial loss of some cell-surface receptors following enzymatic digestion, in line with previous work in peripheral immunology (Supplementary Fig. 1d–f and Supplementary Note 1)3,5–9.”
From Dissection of artifactual and confounding glial signatures by single-cell sequencing of mouse and human brain (2022) · cites this paper as [8]
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- Authors
- Schreurs, R. R. C. E., Drewniak, A., Bakx, R., Corpeleijn, W. E., Geijtenbeek, T. H. B., van Goudoever, J. B., & Bunders, M. J.
- Affiliations
- Amsterdam UMC Location University of Amsterdam (NL)
- University of Amsterdam (NL)
- Corresponding author
- Bunders, M.J.
- Venue
- Journal of Immunological Methods
- Publisher
- Elsevier BV
- Published
- 2017-06
- Volume
- 445
- Pages
- 45-52
- ISSN
- 0022-1759 (ISSN-L)
- OpenAlex
- W2592431882
- Cited by
- 25
- References
- 24
- Citations by year
- 2026: 1 · 2024: 6 · 2023: 3 · 2022: 3 · 2021: 3 · 2020: 4 · 2019: 2 · 2018: 2 · 2017: 1
- FWCI
- 1.63
- Type
- journal article
- Language
- en
- Open access
- hybrid
- doi.org
- Locations
- Journal of Immunological Methods (publishedVersion, cc-by)
- PubMed (publishedVersion)
- Amsterdam UMC (VU Amsterdam) - Institutional Repository (publishedVersion)
- Data Archiving and Networked Services (DANS) (submittedVersion)
- Pure Amsterdam UMC (submittedVersion)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Inflammatory Bowel Disease
- Life Sciences › Immunology and Microbiology › Immunology › T-cell and B-cell Immunology
- Life Sciences › Immunology and Microbiology › Immunology › Immunotoxicology and immune responses
- Keywords
- human intestine, mechanical tissue dissociation, cell yield, cell viability, surface molecule expression, CD4
- MeSH terms
- Flow Cytometry, Humans, Intestinal Mucosa, Intestines, Leukocytes, Mononuclear, Thermolysin, Collagenases
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