CHARACTERIZATION AND FUNCTIONALITY OF HUMAN SERTOLI CELLS

CHUI KITTY; TRIVEDI ALPA; CHERBAVAZ DIANA; DAZIN PAUL; HEMENDINGER RICHELLE; THUY HUYNH AI LAM; MITCHELL JAMES; RABINOVICH GABRIEL; NOBLE-HAEUSSLEIN LINDA; JOHN CONSTANCE

CELL TRANSPLANTATION

COGNIZANT COMMUNICATION CORP

Lugar: MIAMI; Año: 2010

0963-6897

It has long been thought that mammalian Sertoli cells are terminally differentiated and non-dividing post-puberty.  For most in vitro studies testes from immature animals have been the source of Sertoli cells, but these have shown little proliferative ability when cultured.  We isolated and characterized Sertoli cells from human cadaveric testes from seven donors ranging from 12-36 years of age. The cells proliferated readily with a doubling time of approximately four days.  Nuclear EdU incorporation confirmed that dividing cells were the majority of the population. Classical Sertoli cell ultrastructural features, lipid droplet accumulation, expression of GATA4, SOX9, and follicle stimulating hormone receptor (FSHr) were observed by electron and light microscopy. FACS revealed the expression of GATA4 and SOX9 by more than 90% of the cells, and lack of expression of hematopoietic markers. Low detection of endogenous alkaline phosphatase activity after passaging showed that there were few peritubular cells. GATA4 and SOX9 expression was confirmed by RT-PCR, along with stem cell factor (SCF), glial cell line-derived neurotrophic factor (GDNF), and bone morphogenic protein 4 (BMP4). Tight junctions were formed by Sertoli cells plated on transwell inserts coated with fibronectin as revealed by increased transepithelial electrical resistance and polarized secretion of the immunoregulatory protein, Galectin-1.  The ability to propagate functional human Sertoli cells in vitro may facilitate research in testicular cancers, infertility, reproductive toxicology, and spermatogenesis, and potentially could enable therapeutic applications in cell therapy.