Banking of fresh-frozen prostate tissue: methods, validation and use

BJU Int. 2003 Mar;91(4):315-23; discussion 323-4. doi: 10.1046/j.1464-410x.2003.03041.x.

Abstract

Objective: To describe the establishment, methods, validation and use of a bank of fresh-frozen human prostate tissue.

Materials and methods: On obtaining informed patient consent, protocols were followed for banking prostate tissue from any type of prostatectomy or cystoprostatectomy. A pseudobanking procedure was devised to determine the accuracy of assessing the histopathological status of the banked tissue. RNA was extracted, its quality assessed and used for quantitative real-time reverse transcription-polymerase chain reaction for the serine protease hepsin.

Results: To date prostate tissue from 112 patients has been banked, with pseudobanking in 58. The histopathological assessment showed pseudobanked tissue matched adjacent unbanked tissue in 98% of cases for benign vs malignant diagnoses, and in 92% of carcinomas for the Gleason score. Hepsin expression was significantly higher in malignant than in benign tissues (P < 0.0001).

Conclusion: We established a validated method for banking human fresh-frozen prostate tissue and applied it successfully. Hepsin expression can be used to differentiate malignant and benign prostate tissue, and as an indicator of tissue heterogeneity.

Publication types

  • Research Support, Non-U.S. Gov't
  • Validation Study

MeSH terms

  • Cryopreservation / methods*
  • Gene Expression
  • Humans
  • Immunohistochemistry / methods
  • Immunohistochemistry / standards
  • Informed Consent
  • Male
  • Prostate*
  • Prostatectomy
  • Prostatic Diseases / pathology*
  • Quality Control
  • RNA / analysis
  • RNA, Messenger / analysis
  • Reverse Transcriptase Polymerase Chain Reaction / methods
  • Serine Endopeptidases / analysis
  • Tissue Banks / organization & administration
  • Tissue Banks / standards*
  • Tissue Banks / statistics & numerical data

Substances

  • RNA, Messenger
  • RNA
  • Serine Endopeptidases
  • hepsin