Primary culture of microvascular endothelial cells from human benign prostatic hyperplasia.
BACKGROUND
Prostate growth seems to be influenced by paracrine factors like IL-6 originating from the microvascular endothelium. Therefore, our efforts were focused on the primary culture and behavior of microvascular endothelial cells (HPEC) derived from tissue of human benign prostatic hyperplasia (BPH). Until now, the isolation and culture of HPEC from BPH have not been reported.


METHODS
BPH tissue was cut into small cubes and gently squeezed after incubation with dispase. HPEC were cultured from the resulting cell suspension after a stepwise selection by use of superparamagnetic beads coated with antibodies against endothelial specific antigens. HPEC were characterized by flow cytometry and immunohistochemistry. gamma-Glutamyl transpeptidase activity (specific for microvascular endothelium) was measured after dissolution of the HPEC with Triton X-100. After the incubation of HPEC either with ATP, VEGF, or TNF-alpha, the release of IL-6 was measured by enzyme linked immunosorbent assay (ELISA).


RESULTS
HPEC showed a typical endothelial morphology. They were positive for von Willebrand factor, CD31, CD62E (after stimulation with TNF-alpha), alpha-actin and were negative for fibroblastic antigens and PSA. Proliferation was stimulated by vascular endothelial growth factor (VEGF). gamma-Glutamyl transpeptidase activity in HPEC was 6.3 microIU/microg protein, whereas in human umbilical vein endothelial cells (HUVEC) no gamma-glutamyl transpeptidase activity was detectable. The IL-6 secretion of HPEC was stimulated by VEGF and TNF-alpha, but not by ATP and bradykinin.


CONCLUSIONS
For the first time, the primary culture of microvascular endothelial cells from BPH tissue was successfully performed. Our results suggest that HPEC may be actively involved in prostate growth, due to the secretion of regulatory factors such as IL-6.
Stachon
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Institute of Clinical Chemistry, Transfusion, and Laboratory Medicine, University Clinic Bergmannsheil, Ruhr University of Bochum, Bochum, Germany. Axel.Stachon@ruhr-uni-bochum.de
Schlüter
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Köller
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Weisser
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Krieg
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D018929:Cell Culture Techniques / Q000379:methods*; D016228:Endothelial Growth Factors / Q000494:pharmacology; D004727:Endothelium / Q000166:cytology* / Q000502:physiology; D006801:Humans; D007150:Immunohistochemistry; D015850:Interleukin-6 / Q000494:pharmacology; D008222:Lymphokines / Q000494:pharmacology; D008297:Male; D008833:Microcirculation; D011470:Prostatic Hyperplasia / Q000473:pathology*; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology; D042461:Vascular Endothelial Growth Factor A; D042442:Vascular Endothelial Growth Factors
D016228:Endothelial Growth Factors; D015850:Interleukin-6; D008222:Lymphokines; D014409:Tumor Necrosis Factor-alpha; D042461:Vascular Endothelial Growth Factor A; D042442:Vascular Endothelial Growth Factors