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Originally published online as doi:10.2353/jmoldx.2009.080124 on May 21, 2009

Published online before print May 21, 2009
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Journal of Molecular Diagnostics 2009, Vol. 11, No. 4
Copyright © 2009 American Society for Investigative Pathology & Association for Molecular Pathology
DOI: 10.2353/jmoldx.2009.080124

Technical Advances

A Tumor Sorting Protocol that Enables Enrichment of Pancreatic Adenocarcinoma Cells and Facilitation of Genetic Analyses

Zachary S. Boyd*, Rajiv Raja*, Stephanie Johnson{dagger}, David A. Eberhard{ddagger} and Mark R. Lackner*

From the Departments of Development Oncology Diagnostics, * Sample Repository Group, {dagger} and Research Pathology, {ddagger} Genentech, Incorporated, South San Francisco, California

Molecular profiling of human cancer is complicated by both stromal contamination and cellular heterogeneity within samples from tumor biopsies. In this study, we developed a tissue-processing protocol using mechanical dissociation and flow cytometric sorting that resulted in the respective enrichment of stromal and tumor fractions from frozen pancreatic adenocarcinoma samples. Molecular profiling of DNA from the sorted populations using high-density single nucleotide polymorphism arrays revealed widespread chromosomal loss of heterozygosity in tumor fractions but not in either the stromal fraction or unsorted tissue specimens from the same sample. Similarly, a combination of KRAS mutations and chromosomal copy number changes at key pancreatic cancer loci, such as CDK2NA and TP53, was detected in a substantial proportion of the tumor fractions but not in matched stromal fractions from the same sample. This approach to tissue processing could greatly expand the amount of archived tissue that is available for molecular profiling of human cancer and enable a more accurate diagnosis of genetic alterations in patient samples.






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Copyright © 2009 by the American Society for Investigative Pathology and the Association for Molecular Pathology.