ArticleCellular and molecular bioengineering2021
Optimization of a Clinically Relevant Chemical-Mechanical Tissue Dissociation Workflow for Single-Cell Analysis.
Article in Cellular and molecular bioengineering, 2021. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 4 papers.
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Who cites it
4 citing papers in PubMed.
- Recent advancements in tissue dissociation techniques for cell manufacturing single-cell analysis and downstream processing.Stem cells translational medicine · 2025Review
- Electric Field-Based Tissue Dissociation: A Paradigm Shift Toward Scalable, Enzyme-Free Single-Cell Preparation.Cellular and molecular bioengineering · 2025Article
- Innovative Method for Fully Automated, Enzyme-Free Tissue Dissociation and Preparation for Single-Cell Analysis.Cellular and molecular bioengineering · 2025Article
- Lung cancer organoid-based drug evaluation models and new drug development application trends.Translational lung cancer research · 2024Review
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Authors and funding
3 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
introductionWhile single-cell analysis technology has flourished, obtaining single cells from complex tissues continues to be a challenge. Current methods require multiple steps and several hours of processing. This study investigates chemical and mechanical methods for clinically relevant preparation of single-cell suspension from frozen biopsy cores of complex tissues. The developed protocol can be completed in 15 min.
methodsFrozen bovine liver biopsy cores were normalized by weight, dimension, and calculated cellular composition. Various chemical reagents were tested for their capability to dissociate the tissue via confocal microscopy, hemocytometry and quantitative flow cytometry. Images were processed using ImageJ. Quantitative flow cytometry with gating analysis was also used for the analysis of dissociation. Physical modeling simulations were conducted in COMSOL Multiphysics.
resultsA rapid method for tissue dissociation was developed for single-cell analysis techniques. The results of this study show that a combination of 1% type-1 collagenase and pronase or hyaluronidase in 100 U/µL HBSS solution is the most effective at dissociating 2.5 mm thawed bovine liver biopsy cores in 15 min, with dissociation efficiency of 37-42% and viability >90% as verified using live MDA-MB-231 cancer cells. Cellular dissociation is significantly improved by adding a controlled mechanical force during the chemical process, to dissociate 93
conclusionsUnderstanding cellular dissociation in SUPPLEMENTARY INFORMATION: The online version of this article (10.1007/s12195-021-00667-y) contains supplementary material, which is available to authorized users.
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