Phenotypic characterization and functional analysis of human tumor immune infiltration after mechanical and enzymatic disaggregation
Abstract
Multi-parametric flow cytometry analysis is a reliable method for phenotypic and functional characterization of tumor infiltrating immune cells (TIIC). The isolation of infiltrating leukocytes from solid tumors can be achieved through various methods which can be both enzymatic and mechanical; however, these methods may alter cell biology. The aim of this study was to compare the effects of three tissue disaggregation techniques on TIIC biology in breast, kidney and lung tumor specimens. We therefore compared two enzymatic treatments using either collagenase type IA alone or in combination with collagenase type IV and DNase I type II, and one mechanical system (Medimachine™). We evaluated the impact of treatments on cell viability, surface marker integrity and proliferative capacity. We show that cell viability was not significantly altered by treatments. However, enzymatic treatments decreased cell proliferation; specifically collagenases and DNase provoked a significant decrease in detection of surface markers such as CD4, CD8, CD45RA and CD14, indicating that results of phenotypic studies employing these techniques could be affected. In conclusion, mechanical tissue disaggregation by Medimachine™ appears to be optimal to maintain phenotypic and functional TIIC features.
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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 [5]
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- Authors
- Grange, C., Létourneau, J., Forget, M.-A., Godin-Ethier, J., Martin, J., Liberman, M., Latour, M., Widmer, H., Lattouf, J.-B., Piccirillo, C. A., Cailhier, J.-F., & Lapointe, R.
- Affiliations
- Hôpital Notre-Dame (CA)
- Centre Hospitalier de l’Université de Montréal (CA)
- McGill University Health Centre (CA)
- Corresponding author
- Lapointe, Réjean
- ORCID
- Venue
- Journal of Immunological Methods
- Publisher
- Elsevier BV
- Published
- 2011-09
- Volume
- 372
- Issue
- 1-2
- Pages
- 119-126
- ISSN
- 0022-1759 (ISSN-L)
- OpenAlex
- W2010681714
- Cited by
- 29
- References
- 27
- Citations by year
- 2026: 1 · 2025: 2 · 2024: 1 · 2023: 2 · 2022: 2 · 2021: 2 · 2020: 4 · 2019: 3 · 2018: 1 · 2017: 2 · 2016: 1 · 2015: 1 · 2014: 3 · 2013: 1 · 2012: 3
- FWCI
- 1.10
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Journal of Immunological Methods (publishedVersion)
- PubMed (publishedVersion)
- Funders
- Université de Montréal
- Topics
- Health Sciences › Medicine › Oncology › Cancer Immunotherapy and Biomarkers
- Life Sciences › Immunology and Microbiology › Immunology › Immune cells in cancer
- Life Sciences › Immunology and Microbiology › Immunology › T-cell and B-cell Immunology
- Keywords
- tumor-infiltrating immune cells, tissue disaggregation, mechanical tissue dissociation, enzymatic tissue dissociation, type IV collagenase, DNase I, cell viability, surface marker expression, cell proliferation, breast tumors, kidney tumors, lung cancer, tumor immune infiltration
- MeSH terms
- Breast Neoplasms, Cell Separation, Female, Flow Cytometry, Humans, Kidney Neoplasms, Lung Neoplasms, Lymphocytes, Tumor-Infiltrating, Cell Proliferation
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