human prostate endothelial cells  (PromoCell)

Journal: Molecular cancer therapeutics

Article Title: Targeted BikDD expression kills androgen-dependent and castration-resistant prostate cancer cells

doi: 10.1158/1535-7163.MCT-13-1004

Figure Lengend Snippet: (A) Schematic diagram of BikDD constructs CMV-BikDD (pUK21-CMV-BikDD), AT-VISA-BikDD (pUK21-AT-VISA-BikDD) and control (ctrl; pUK21-AT-VISA). (B) LNCaP, PC-3, and WI-38 cells transiently transfected with indicated plasmids and then stimulated with R1881 at 1 nM in medium containing charcoal/dextran-treated FBS. After transfection, cell lysates were subjected to Western blotting to detect the expression of BikDD (left). Quantitation is shown on the right. (C) In vitro killing effect of BikDD. A panel of AR+ prostate cancer (LNCaP and LAPC-4), AR- prostate cancer (PC-3 and DU145) and normal (PrEC, WI-38 and HUVEC) cells were transiently co-transfected with 2 μg of CMV-BikDD, AT-VISA-BikDD or control, plus 100 ng of CMV-Luc as indicated and then stimulated with R1881 at 1 nM in medium containing charcoal/dextran-treated FBS. Forty-eight hours post transfection, cells were imaged for 30 seconds by the IVIS™ Imaging System to determine the luciferase reporter activity 5 min following the addition of 5 ng/ml of D-luciferin. Top, representative images. Bottom, quantitation of the signals relative to negative control (set as 100%). The data represent the mean of three independent experiments. Error bars, SD.

Article Snippet: Normal prostate epithelial cells (PrEC) and normal human vascular endothelial cells (HUVEC) were also purchased from ATCC.

Techniques: Construct, Control, Transfection, Western Blot, Expressing, Quantitation Assay, In Vitro, Imaging, Luciferase, Activity Assay, Negative Control

Journal: Oncotarget

Article Title: Isolation and genome-wide expression and methylation characterization of CD31 + cells from normal and malignant human prostate tissue

doi:

Figure Lengend Snippet: Clinically examined and representative samples were mounted in OCT. Prostate tissues obtained from robotic radical prostectomy specimens from each patient were macrodissected for prostatic adenocarcinoma (tumor) and matched histologically benign regions. Portion of freshly macrodissected prostate tissues (tumor and non-tumor) were immediately digested and cultured in endothelial cell selection medium. Given that enough tissues were obtained, a portion of the tissues used for ECs isolation were embedded in OCT for frozen sections. ECs were isolated by CD31 magnetic Dynabead® and further enriched by CD31 fluorescent activated cell sorting (FACS).

Article Snippet: Primary cultures of human prostate endothelial cells (NdECs and TdECs) were cultured in endothelial growth medium [Endothelial Cell Growth Medium MV2 with Supplement Mix (PromoCell GmbH, Heidelberg, Germany) and 100 U/ml penicillin and 100 μg/ml streptomycin (Invitrogen)].

Techniques: Cell Culture, Selection, Isolation, FACS

Journal: Oncotarget

Article Title: Isolation and genome-wide expression and methylation characterization of CD31 + cells from normal and malignant human prostate tissue

doi:

Figure Lengend Snippet: (A) Representative photomicrographs of hematoxylin and eosin stained frozen sections of benign (left) and malignant (right) prostatic tissues of macrodissected tissues used for ECs isolation. ECs are highlighted in the frozen sections by CD31 immunostaining (Original magnification, x200). (B) Representative immunofluorescent photomicrographs of CD31 (green) and vWF (red) expression and uptake of DiI-Ac-LDL (red) (original magnification, ×200) in CD31 + prostate TdECs and NdECs. Absence of pan-cytokeratin expression by immunofluorescence was observed in CD31 + prostate TdECs and NdECs. LNCaP was used as positive control for pan-cytokeratin immunofluorescence. Nuclei are stained with DAPI (blue). Endothelial tube network was formed by primary cultures of prostate TdECs and NdECs (original magnification, x100). (C) Representative reverse transcription-PCR analysis of RNA from primary cultures of TdECs and NdECs, HUVECs, or LNCaP cells using human-specific primers for human CD31, CD34, ICAM-1, VCAM-1, VEGFR1, VEGFR2, AR and PSA. GAPDH was used as a loading control.

Article Snippet: Primary cultures of human prostate endothelial cells (NdECs and TdECs) were cultured in endothelial growth medium [Endothelial Cell Growth Medium MV2 with Supplement Mix (PromoCell GmbH, Heidelberg, Germany) and 100 U/ml penicillin and 100 μg/ml streptomycin (Invitrogen)].

Techniques: Staining, Isolation, Immunostaining, Expressing, Immunofluorescence, Positive Control, Reverse Transcription, Control

Journal: Oncotarget

Article Title: Isolation and genome-wide expression and methylation characterization of CD31 + cells from normal and malignant human prostate tissue

doi:

Figure Lengend Snippet: Quantitative real-time PCR was used to validate gene expression of AREG , JMY , EPB41 , GMNN and FAM53C in endothelial cells derived from malignant lesions vs benign lesions in AA and CA patients with prostate cancer. Relative gene expression level for qRT-PCR was normalized to the reference gene GAPDH . Gene expression from microarray was plotted together with the qRT-PCR results. Results were shown as Mean ± SD (triplicate). “*” represents p value <0.05 by t-Test.

Article Snippet: Primary cultures of human prostate endothelial cells (NdECs and TdECs) were cultured in endothelial growth medium [Endothelial Cell Growth Medium MV2 with Supplement Mix (PromoCell GmbH, Heidelberg, Germany) and 100 U/ml penicillin and 100 μg/ml streptomycin (Invitrogen)].

Techniques: Real-time Polymerase Chain Reaction, Gene Expression, Derivative Assay, Quantitative RT-PCR, Microarray