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Preferential loss of abnormal prostate carcinoma cells by collagenase treatment
J J König1, J W van Dongen, F H Schröder
1Department of Urology, Erasmus University, Rotterdam, The Netherlands.
Abstract:
Effects of two different methods of tumor disaggregation on flow cytometric ploidy distribution and intact cell yield were investigated. Either mechanical disaggregation or collagenase digestion was applied to 35 prostate tumor specimens. Seven collagenase-treated samples failed to yield any intact cells, whereas with mechanical disaggregation in all cases a sufficient number of intact cells were obtained. No differences in the FCM ploidy distribution of tumors with a DNA diploid stemline were observed comparing both techniques. In DNA aneuploid tumors, however, collagenase treatment had an adverse effect on the abnormal cell populations. In 14/17 of such tumors, the abnormal cell populations were significantly reduced; in eight of these the percentage of DNA aneuploid cells declined even below the minimum percentage (10%) that was defined for DNA aneuploidy. Since collagenase is a widely used enzyme for tissue disaggregation, especially in tumor cytogenetics, the presented data will have consequences for the interpretation of results obtained by methods involving the use of this enzyme.
Insights
Mechanical tumor disaggregation is superior to collagenase digestion for preserving intact cells and accurately assessing DNA aneuploidy in prostate tumors. Collagenase treatment significantly reduces abnormal cell populations, impacting diagnostic interpretations.
Area of Science:
- Oncology
- Cell Biology
- Biotechnology
Background:
- Accurate tumor cell analysis is crucial for diagnosis and treatment planning.
- Tissue disaggregation methods can impact cell yield and ploidy distribution.
- Collagenase is commonly used for tissue digestion in research.
Purpose of the Study:
- To compare mechanical disaggregation and collagenase digestion for prostate tumor samples.
- To evaluate the impact of these methods on intact cell yield and flow cytometric ploidy distribution.
- To determine the consequences for interpreting flow cytometry (FCM) data.
Main Methods:
- 35 prostate tumor specimens were processed using either mechanical disaggregation or collagenase digestion.
- Intact cell yield was assessed for both methods.
- Flow cytometric analysis was performed to determine DNA ploidy distribution.
Main Results:
- Mechanical disaggregation consistently yielded sufficient intact cells, while 7 collagenase-treated samples failed.
- No differences in ploidy distribution were observed for diploid tumors.
- Collagenase treatment significantly reduced abnormal cell populations in aneuploid tumors, with 8 cases falling below the 10% threshold.
Conclusions:
- Mechanical disaggregation is preferable for obtaining intact cells and reliable ploidy analysis in prostate tumors.
- Collagenase digestion can artifactually reduce the percentage of aneuploid cells, potentially leading to misdiagnosis.
- Results highlight the need to consider disaggregation methods when interpreting FCM ploidy data in tumor analysis.