Obtaining Undifferentiated Cells From Testis Biopsy

Azoospermia Clinical Trial

Official title:

Isolation, Culture and Characterization Stem Cells From Testis Biopsy

Clinical Trial Details — Status: Completed

Administrative data

• NCT number: NCT01375062
• Other study ID #: 0902-057-F-CS
• Status: Completed
• Phase: N/A
• First received: June 7, 2011
• Last updated: October 16, 2015
• Start date: May 2009
• Est. completion date: July 2011

Study information

• Verified date: March 2014
• Source: Igenomix
• Contact: n/a
• Is FDA regulated: No
• Health authority: Spain: Ministry of Health
• Study type: Interventional

Clinical Trial Summary

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

Description:

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).

Recruitment information / eligibility

• Status: Completed
• Enrollment: 65
• Est. completion date: July 2011
• Est. primary completion date: July 2011
• Accepts healthy volunteers: Accepts Healthy Volunteers
• Gender: Male
• Age group: 18 Years and older

Eligibility

Inclusion Criteria:

-azoospermic patients

Study Design

Intervention Model: Single Group Assignment, Masking: Open Label, Primary Purpose: Treatment

Related Conditions & MeSH terms

• Azoospermia

Intervention

Other:
• analyse and proliferate the cells
Obtention of excess tissue from testicular biopsies and maintained in 4ºC until procedures. Digestion of the biopsies through controlled enzymes Replication of the cells Selection undifferentiated cells Expansion of the purification cells population Characterization of the cell population Assessment of pluripotent and germinal cell potential. Possible maturation of the cells.

Locations

Country	Name	City	State
Spain	IVI Valencia	Valencia

Sponsors (1)

Lead Sponsor	Collaborator
Igenomix

Country where clinical trial is conducted

Spain, 

Outcome

Type	Measure	Description	Time frame	Safety issue
Primary	Assessment of potential germinal and pluripotent of the isolated cultures.	•The spermatogonial cells isolated from testicular biopsies of azoospermic patients will be matured in vitro for thegeneration of sperm for infertility treatment.; * Undifferentiated cells of the testis biopsy will be characterized as degrees of plasticity for proof that they are capable of regeneration spermatogenesis.	one month	No