Clinical Trial: Obtaining Undifferentiated Cells From Testis Biopsy

Study Status: Completed
Recruit Status: Completed
Study Type: Interventional

Official Title: Isolation, Culture and Characterization Stem Cells From Testis Biopsy

Brief Summary: The purpose of this study is to isolate and culture stem cell from adult human testicular biopsies for future transplantation in fertility preservation.

Detailed Summary: Testicular biopsies were obtained from azoospermic patients as part of their diagnosis and work-up after obtaining a signed informed consent. A portion of each sample was mechanically dissected in PBS with 1% Pen/Strep. For enzymatic digestion samples were incubated with 0.5g/ml Collagenase type 1A for 20min and TripLE Select for 15min at 37ºC on a shaker and then filtered with 30µm and centrifuged at 1,000rpm. The pellets were seeded and incubated in embryonic stem cell media (hESC) with glial cell line-derived neurotrophic factor (GDNF 4ng/mL) for 5-7 days at 37ºC (1st culture). To identify and isolate SSCs they were marked with CD49f antibody and FACS sorted. Positive cells were cultured in gelatine plates containing hESC media with leukaemia inhibitory factor (LIF1000U/mL) (2nd culture).
Sponsor: Igenomix

Current Primary Outcome: Assessment of potential germinal and pluripotent of the isolated cultures. [ Time Frame: one month ]

Original Primary Outcome: Same as current

Current Secondary Outcome:

Original Secondary Outcome:

Information By: Igenomix

Dates:
Date Received: June 7, 2011
Date Started: May 2009
Date Completion:
Last Updated: October 16, 2015
Last Verified: March 2014