World J Mens Health.  2020 Jan;38(1):123-131. 10.5534/wjmh.180091.

A Simple and Nonenzymatic Method to Isolate Human Corpus Cavernosum Endothelial Cells and Pericytes for the Study of Erectile Dysfunction

Affiliations
  • 1National Research Center for Sexual Medicine and Department of Urology, Inha University School of Medicine, Incheon, Korea. jksuh@inha.ac.kr rjk0929@inha.ac.kr
  • 2Department of Urology, Yantai Yuhuangding Hospital Affiliated to Medical College of Qingdao University, Yantai, China.

Abstract

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.

Keyword

Diabetes mellitus; Endothelial cell; Erectile dysfunction; Pericytes

MeSH Terms

Blotting, Western
Cells, Cultured
Diabetes Mellitus
Endothelial Cells*
Erectile Dysfunction*
Gravitation
Humans*
Immunohistochemistry
Male
Methods*
Pericytes*
Phenotype
Platelet-Derived Growth Factor
von Willebrand Factor
Platelet-Derived Growth Factor
von Willebrand Factor
Full Text Links
  • WJMH
Actions
Cited
CITED
export Copy
Close
Share
  • Twitter
  • Facebook
Similar articles
    DB Error: unknown error