Rapid and Short-term Extracellular Matrix-mediated In Vitro Culturing of Tumor and Nontumor Human Primary Prostate Cells From Fresh Radical Prostatectomy Tissue
| Autor: | Vladimir Mouraviev, David M. Albala, Ashok C. Chander, Brad J. Hogan, Grannum R. Sant, Jonathan S. Varsanik, Michael S. Manak, Stephen M. Zappala |
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| Rok vydání: | 2017 |
| Předmět: |
Male
0301 basic medicine Pathology medicine.medical_specialty Time Factors Cell Survival Urology medicine.medical_treatment Cell Culture Techniques Cell Growth Processes Immunofluorescence Focal adhesion Extracellular matrix 03 medical and health sciences 0302 clinical medicine Prostate Biopsy Cell Adhesion Tumor Cells Cultured medicine Humans Prostatectomy medicine.diagnostic_test business.industry Kinase Biopsy Needle Prostatic Neoplasms Epithelial Cells In vitro Extracellular Matrix 030104 developmental biology medicine.anatomical_structure 030220 oncology & carcinogenesis Cancer research Stromal Cells business |
| Zdroj: | Urology. 105:91-100 |
| ISSN: | 0090-4295 |
| DOI: | 10.1016/j.urology.2017.03.029 |
| Popis: | Objective To culture prostate cells from fresh biopsy core samples from radical prostatectomy (RP) tissue. Further, given the genetic heterogeneity of prostate cells, the ability to culture single cells from primary prostate tissue may be of importance toward enabling single-cell characterization of primary prostate tissue via molecular and cellular phenotypic biomarkers. Methods A total of 260 consecutive tissue samples from RPs were collected between October 2014 and January 2016, transported at 4°C in serum-free media to an off-site central laboratory, dissociated, and cultured. A culture protocol, including a proprietary extracellular matrix formulation (ECMf), was developed that supports rapid and short-term single-cell culture of primary human prostate cells derived from fresh RP samples. Results A total of 251 samples, derived from RP samples, yielded primary human tumor and nontumor prostate cells. Cultured cells on ECMf exhibit (1) survival after transport from the operating room to the off-site centralized laboratory, (2) robust (>80%) adhesion and survival, and (3) expression of different cell-type-specific markers. Cells derived from samples of increasing Gleason score exhibited a greater number of focal adhesions and more focal adhesion activation as measured by phospho-focal adhesion kinase (Y397) immunofluorescence when patient-derived cells were cultured on ECMf. Increased Ki67 immunofluorescence levels were observed in cells derived from cancerous RP tissue when compared to noncancerous RP tissue. Conclusion By utilizing a unique and defined extracellular matrix protein formulation, tumor and nontumor cells derived from primary human prostate tissue can be rapidly cultured and analyzed within 72 hours after harvesting from RP tissue. |
| Databáze: | OpenAIRE |
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