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Novel method to characterize immune cells from human prostate tissue

BACKGROUND Benign prostatic hyperplasia (BPH) is the most common benign adenoma and prostate cancer is the most frequent malignancy in men over 50 years of age in the Western world, where it remains a significant health problem. Prostate lesions are known to contain immune cells, which may contribut...

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Published in:The Prostate 2014-10, Vol.74 (14), p.1391-1399
Main Authors: Norström, Melissa M., Rådestad, Emelie, Stikvoort, Arwen, Egevad, Lars, Bergqvist, Mats, Henningsohn, Lars, Mattsson, Jonas, Levitsky, Victor, Uhlin, Michael
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container_end_page 1399
container_issue 14
container_start_page 1391
container_title The Prostate
container_volume 74
creator Norström, Melissa M.
Rådestad, Emelie
Stikvoort, Arwen
Egevad, Lars
Bergqvist, Mats
Henningsohn, Lars
Mattsson, Jonas
Levitsky, Victor
Uhlin, Michael
description BACKGROUND Benign prostatic hyperplasia (BPH) is the most common benign adenoma and prostate cancer is the most frequent malignancy in men over 50 years of age in the Western world, where it remains a significant health problem. Prostate lesions are known to contain immune cells, which may contribute to the immune control of tumor progression. However, due to their low numbers and restricted access to necessary material it is difficult to isolate immune cells from prostate tissue to characterize their immunological features. METHODS An efficient and robust method was developed to process prostate tissue and isolate immune cells for phenotypic analysis by multicolor flow cytometry as downstream application. Fresh prostate tissue from 11 patients undergoing surgery for bladder outlet obstruction due to BPH was processed to evaluate the number, viability, yield, and frequency of various immune cell types. RESULTS The presented method does not include enzymatic digestion nor incubation steps at 37°C, increasing cellular viability and avoiding possible phenotypic modification. Various immune cell populations were detected in all patient samples and the median cellular viability was 90%. The number of detected events of individual cell populations varied between patients. The median frequency of different immune cell populations also varied, being 87% for the CD3‐ and 15% for the CD3+ cell population. CONCLUSIONS This novel method will allow the phenotypic characterization of immune cell populations present in tumor tissue of prostate cancer patients and promote development of novel approaches to immunotherapy of the disease. Prostate 74:1391–1399, 2014. © 2014 Wiley Periodicals, Inc.
doi_str_mv 10.1002/pros.22854
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Prostate lesions are known to contain immune cells, which may contribute to the immune control of tumor progression. However, due to their low numbers and restricted access to necessary material it is difficult to isolate immune cells from prostate tissue to characterize their immunological features. METHODS An efficient and robust method was developed to process prostate tissue and isolate immune cells for phenotypic analysis by multicolor flow cytometry as downstream application. Fresh prostate tissue from 11 patients undergoing surgery for bladder outlet obstruction due to BPH was processed to evaluate the number, viability, yield, and frequency of various immune cell types. RESULTS The presented method does not include enzymatic digestion nor incubation steps at 37°C, increasing cellular viability and avoiding possible phenotypic modification. Various immune cell populations were detected in all patient samples and the median cellular viability was 90%. The number of detected events of individual cell populations varied between patients. The median frequency of different immune cell populations also varied, being 87% for the CD3‐ and 15% for the CD3+ cell population. CONCLUSIONS This novel method will allow the phenotypic characterization of immune cell populations present in tumor tissue of prostate cancer patients and promote development of novel approaches to immunotherapy of the disease. 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The number of detected events of individual cell populations varied between patients. The median frequency of different immune cell populations also varied, being 87% for the CD3‐ and 15% for the CD3+ cell population. CONCLUSIONS This novel method will allow the phenotypic characterization of immune cell populations present in tumor tissue of prostate cancer patients and promote development of novel approaches to immunotherapy of the disease. 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subjects Aged
Aged, 80 and over
cell isolation
Flow Cytometry - methods
Humans
immune cell populations
Immunophenotyping
Male
Medicin och hälsovetenskap
Middle Aged
multicolor flow cytometry
prostate tissue
Prostatic Neoplasms - immunology
Prostatic Neoplasms - pathology
single cell suspension
title Novel method to characterize immune cells from human prostate tissue
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