A Simple and Nonenzymatic Method to Isolate Human Corpus Cavernosum Endothelial Cells and Pericytes for the Study of Erectile Dysfunction
| Autor: | Jiyeon Ock, Min Ji Choi, Ji Kan Ryu, Do Hwan Seong, Kalyan Ghatak, Hai Rong Jin, Mi-Hye Kwon, Nguyen Nhat Minh, Guo Nan Yin, Kang-Moon Song, Jun Kyu Suh |
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| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Předmět: |
Aging
Pathology medicine.medical_specialty erectile dysfunction Urology medicine.medical_treatment Immunocytochemistry 030232 urology & nephrology lcsh:Medicine lcsh:RC870-923 pericytes 03 medical and health sciences 0302 clinical medicine Von Willebrand factor Western blot medicine Pharmacology (medical) Tube formation Matrigel 030219 obstetrics & reproductive medicine biology medicine.diagnostic_test business.industry Health Policy Growth factor lcsh:R Public Health Environmental and Occupational Health lcsh:Diseases of the genitourinary system. Urology Endothelial stem cell Psychiatry and Mental health medicine.anatomical_structure Reproductive Medicine diabetes mellitus biology.protein endothelial cell Original Article Pericyte business Male Sexual Health and Dysfunction |
| Zdroj: | The World Journal of Men's Health, Vol 38, Iss 1, Pp 123-131 (2020) The World Journal of Men's Health |
| ISSN: | 2287-4690 2287-4208 |
| Popis: | Purpose To establish a simple and nonenzymatic technique to isolate endothelial cells (ECs) and pericytes from human corpus cavernosum tissue and to evaluate the angiogenic ability of the human cavernous EC or pericytes for the study of high glucose-induced angiopathy. Materials and methods For primary human cavernous EC culture, cavernous tissues were implanted into Matrigel in dishes. For primary human cavernous pericyte culture, cavernous tissues were settled by gravity into dishes. We performed immunocytochemistry and Western blot to determine phenotype and morphologic changes from passage 1 to 5. The primary cultured cells were exposed to a normal-glucose (5 mmol/L) or a high-glucose (30 mmol/L) condition, and then tube formation assay was done. Results We successfully isolated high-purity EC and pericytes from human corpus cavernosum tissue. Primary cultured EC showed highly positive staining for von Willebrand factor, and pericyte revealed positive staining for NG2 and platelet-derived growth factor receptor-β. Primary cultured EC and pericytes maintained their cellular characteristics up to passage 2 or 3. However, we observed significant changes in their typical phenotype from the passage 4 and morphological characteristics from the passage 3. Human cavernous EC or pericytes formed well-organized capillary-like structures in normal-glucose condition, whereas severely impaired tube formation was detected in high-glucose condition. Conclusions This study provides a simple and nonenzymatic method for primary culture of human cavernous EC and pericytes. Our study will aid us to understand the pathophysiology of diabetic erectile dysfunction, and also be a valuable tool for determining the efficacy of candidate therapeutic targets. |
| Databáze: | OpenAIRE |
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