Collagenase and Dispase enzymes disrupt lymphocyte surface molecules
Abstract
Collagenase and Dispase enzymes are often used to disaggregate tissue. In this study, we examined by flow cytometry the effects of these enzyme preparations on peripheral blood lymphocyte surface marker expression. Peripheral blood mononuclear cells (PBMC) were obtained from seven healthy volunteers. Cells were incubated with collagenase (128 U/ml, type 1 A) for 3 h and overnight and with Dispase (1.6 mg/ml, grade II) for 15 min, 30 min, 1 h, 3 h and overnight. The intensity of expression of CD3, CD4, CD8, alpha beta and gamma delta T cell receptors was decreased by 25-40% in all cases, while CD4+ and CD8+ lymphocyte populations were undetectable after treatment with Dispase. Moreover, reappearance of these surface molecules did not occur following the incubation of cells in culture medium for 3 h. The results of this study emphasise the importance of considering the influence of isolation procedures on cell characteristics.
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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 [3]
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Gao, C., Hollingsworth, R. E., & Hurt, E. M. (2012) · Springer eBooks · doi:10.1007/978-94-007-4590-2_1
Nicol, M. R. (2019) · Carolina Digital Repository (University of North Carolina at Chapel Hill) · doi:10.17615/x9b2-e491
kim, Y., Yun, H., Noh, S., Lim, S., Chung, J. Y., Park, J.-Y., & Park, D. Y. (2026) · Tissue Engineering and Regenerative Medicine · doi:10.1007/s13770-026-00805-5
Bartlett, D. C. (2015) · University of Birmingham Institutional Research Archive (University of Birmingham)
Effects of Collagenase on Human Hematopoietic Cells
Jelinek, N., Schmidt, S., Thoma, S. J., Wandrey, C., & Biselli, M. (2008) · Springer eBooks · doi:10.1007/0-306-46875-1_117
- Authors
- Abuzakouk, M., Feighery, C., & O'Farrelly, C.
- Affiliations
- St. James's Hospital (IE)
- Venue
- Journal of Immunological Methods
- Publisher
- Elsevier BV
- Published
- 1996-08
- Volume
- 194
- Issue
- 2
- Pages
- 211-216
- ISSN
- 0022-1759 (ISSN-L)
- OpenAlex
- W2068672112
- Cited by
- 90
- References
- 12
- Citations by year
- 2026: 1 · 2025: 2 · 2024: 4 · 2023: 3 · 2022: 5 · 2021: 4 · 2020: 4 · 2019: 4 · 2018: 5 · 2017: 1 · 2016: 6 · 2015: 6 · 2014: 10 · 2013: 5 · 2012: 2
- FWCI
- 1.37
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Journal of Immunological Methods (publishedVersion)
- PubMed (publishedVersion)
- Topics
- Life Sciences › Immunology and Microbiology › Immunology › T-cell and B-cell Immunology
- Life Sciences › Immunology and Microbiology › Immunology › Immunotoxicology and immune responses
- Health Sciences › Medicine › Radiology, Nuclear Medicine and Imaging › Monoclonal and Polyclonal Antibodies Research
- Keywords
- collagenase, dispase, lymphocyte surface markers, peripheral blood mononuclear cells, T cell receptor expression, CD3, CD4, CD8, γδ T cells
- MeSH terms
- Antigens, Surface, Flow Cytometry, Humans, Endopeptidases, T-Lymphocytes, CD4-Positive T-Lymphocytes, T-Lymphocyte Subsets, Collagenases, CD8-Positive T-Lymphocytes
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