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


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).


CONCLUSIONS
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.
Riddick
A C P
AC
Norfolk and Norwich University Hospital, Norwich, UK.
Barker
C
C
Sheriffs
I
I
Bass
R
R
Ellis
V
V
Sethia
K K
KK
Edwards
D R
DR
Ball
R Y
RY
D015925:Cryopreservation / Q000379:methods*; D015870:Gene Expression; D006801:Humans; D007150:Immunohistochemistry / Q000379:methods / Q000592:standards; D007258:Informed Consent; D008297:Male; D011467:Prostate; D011468:Prostatectomy; D011469:Prostatic Diseases / Q000473:pathology*; D011786:Quality Control; D012313:RNA / Q000032:analysis; D012333:RNA, Messenger / Q000032:analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D012697:Serine Endopeptidases / Q000032:analysis; D014015:Tissue Banks / Q000458:organization & administration / Q000592:standards* / Q000706:statistics & numerical data
D012333:RNA, Messenger; D012313:RNA; D012697:Serine Endopeptidases; C055645:hepsin