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Primary culture of microvascular endothelial cells from human benign prostatic hyperplasia
Author(s) -
Stachon Axel,
Schlüter Tanja,
Köller Manfred,
Weisser Heike,
Krieg Michael
Publication year - 2001
Publication title -
the prostate
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.295
H-Index - 123
eISSN - 1097-0045
pISSN - 0270-4137
DOI - 10.1002/pros.1094
Subject(s) - endothelium , du145 , dispase , cd31 , microbiology and biotechnology , biology , endothelial stem cell , paracrine signalling , pathology , angiogenesis , medicine , endocrinology , prostate , in vitro , cancer research , collagenase , biochemistry , receptor , cancer , lncap , enzyme
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. γ‐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‐α, 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‐α), α‐actin and were negative for fibroblastic antigens and PSA. Proliferation was stimulated by vascular endothelial growth factor (VEGF). γ‐Glutamyl transpeptidase activity in HPEC was 6.3 μIU/μg protein, whereas in human umbilical vein endothelial cells (HUVEC) no γ‐glutamyl transpeptidase activity was detectable. The IL‐6 secretion of HPEC was stimulated by VEGF and TNF‐α, 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. Prostate 48:156–164, 2001. © 2001 Wiley‐Liss, Inc.