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Updated: Apr 21, 2026
Enrichment and Characterization of the Tumor Immune and Non-immune Microenvironments in Established Subcutaneous Murine Tumors
Published on: June 7, 2018
An optimized disaggregation method for human lung tumors that preserves the phenotype and function of the immune
Jon G Quatromoni1, Sunil Singhal1, Pratik Bhojnagarwala1
1Departments of *Surgery and Medicine, University of Pennsylvania School of Medicine, and Department of Pathology and Laboratory Medicine, Children's Hospital of Philadelphia and University of Pennsylvania School of Medicine, Philadelphia, Pennsylvania, USA.
Insights
Preparing human lung tumors for immunology research is crucial. Current methods alter immune cell data, but a new gentle technique preserves cell integrity and surface markers for accurate immunologic studies.
Area of Science:
- Immunology
- Oncology
- Biotechnology
Background:
- Accurate immunologic studies rely on proper human tissue preparation.
- Current human lung tumor disaggregation methods lack rigorous testing for data fidelity.
- Existing techniques can alter immune cell composition and surface molecule expression.
Purpose of the Study:
- To critically evaluate existing human lung tumor preparation techniques for immunologic studies.
- To develop and validate a novel, optimized method for human lung tumor disaggregation.
- To ensure the preservation of immune cell populations and surface marker integrity.
Main Methods:
- Evaluation of current literature-based tissue disaggregation techniques for human lung tumors.
- Development of a novel method combining gentle mechanical manipulation with an optimized enzyme cocktail at low doses.
- Assessment of cell yield, viability, immune cell population retrieval, and surface marker expression.
Main Results:
- Current methods effectively digest cellular attachments and extracellular matrices (ECMs).
- Existing techniques frequently alter immune cell composition and/or surface molecule expression.
- The novel method optimized cell yield and viability, retrieved all major tumor-associated cell populations, and maintained cell-surface marker expression.
Conclusions:
- Established methods for human lung tumor processing can compromise immunologic data integrity.
- A novel, rigorously tested disaggregation method was developed for human lung tumors.
- This new approach ensures accurate immune cell profiling and functional analysis in tumor immunology research.
Abstract:
Careful preparation of human tissues is the cornerstone of obtaining accurate data in immunologic studies. Despite the essential importance of tissue processing in tumor immunology and clinical medicine, current methods of tissue disaggregation have not been rigorously tested for data fidelity. Thus, we critically evaluated the current techniques available in the literature that are used to prepare human lung tumors for immunologic studies. We discovered that these approaches are successful at digesting cellular attachments and ECMs; however, these methods frequently alter the immune cell composition and/or expression of surface molecules. We thus developed a novel approach to prepare human lung tumors for immunologic studies by combining gentle mechanical manipulation with an optimized cocktail of enzymes used at low doses. This enzymatic digestion cocktail optimized cell yield and cell viability, retrieved all major tumor-associated cell populations, and maintained the expression of cell-surface markers for lineage definition and in vivo effector functions. To our knowledge, we present the first rigorously tested disaggregation method designed for human lung tumors.
