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Disaggregation of human solid tumours by combined mechanical and enzymatic methods.
British Journal of Cancer
|January 1, 1985
Summary
Comparing tumor disaggregation methods, combined mechanical and enzymatic techniques yield significantly more viable cells than mechanical methods alone. Long-term trypsinization at 4°C provides the most representative cell yield for downstream analysis.
Area of Science:
- Oncology
- Cell Biology
- Biotechnology
Background:
- Efficiently isolating viable tumor cells is crucial for cancer research and therapeutic development.
- Current methods for tumor disaggregation vary in effectiveness, impacting downstream applications.
- Optimizing cell yield and representativeness is essential for accurate biological analysis.
Purpose of the Study:
- To compare the efficacy of combined mechanical-enzymatic disaggregation techniques versus a purely mechanical method for tumor tissue processing.
- To evaluate different enzymatic disaggregation protocols (long-term cold vs. short-term warm trypsinization).
- To assess the impact of disaggregation methods on viable cell yield, plating efficiency, tumor formation in vivo, and DNA content.
Main Methods:
- Tumor tissues were processed using a simple mechanical disaggregation method.
- Combined methods involved mechanical comminution followed by trypsin incubation.
- Enzymatic protocols included long-term trypsinization at 4°C and repeated short-term trypsinization at 37°C.
- Evaluation metrics included viable cell yield, plating efficiency, tumorogenicity in nude mice, and DNA distribution via flow cytometry.
Main Results:
- Combined mechanical-enzymatic techniques yielded 2-10 x 10^7 viable cells/g of tissue, significantly higher than the mechanical method.
- Long-term trypsinization at 4°C provided a representative cell yield from all tested tumors.
- DNA analysis confirmed the superiority of the long-term trypsinization procedure for representative cell recovery.
Conclusions:
- Combined mechanical and enzymatic tumor disaggregation significantly enhances viable cell recovery compared to mechanical methods alone.
- Long-term, low-temperature trypsinization is the optimal enzymatic method for obtaining representative tumor cell yields for comprehensive analysis.
- The choice of disaggregation technique critically influences the quality and quantity of cells for downstream biological studies.