title: Optimizing cryopreservation of human spermatogonial stem cells: comparing the effectiveness of testicular tissue and single cell suspension cryopreservation.
abstract: OBJECTIVE
  To determine whether optimal human spermatogonial stem cell (SSC) cryopreservation is best achieved with testicular tissue or single cell suspension cryopreservation. This study compares the effectiveness between these two approaches by using testicular SSEA-4+ cells, a known population containing SSCs.
  
  
  METHODS
  In vitro human testicular tissues.
  
  
  METHODS
  Academic research unit.
  
  
  METHODS
  Adult testicular tissues (n=4) collected from subjects with normal spermatogenesis and normal fetal testicular tissues (n=3).
  
  
  METHODS
  Testicular tissue versus single cell suspension cryopreservation.
  
  
  METHODS
  Cell viability, total cell recovery per milligram of tissue, as well as viable and SSEA-4+ cell recovery.
  
  
  RESULTS
  Single cell suspension cryopreservation yielded higher recovery of SSEA-4+ cells enriched in adult SSCs, whereas fetal SSEA-4+ cell recovery was similar between testicular tissue and single cell suspension cryopreservation.
  
  
  CONCLUSIONS
  Adult and fetal human SSEA-4+ populations exhibited differential sensitivity to cryopreservation based on whether they were cryopreserved in situ as testicular tissues or as single cells. Thus, optimal preservation of human SSCs depends on the patient's age, type of samples cryopreserved, and end points of therapeutic applications.
authors.affiliation: Department of Obstetrics, Gynecology and Reproductive Sciences, University of California, San Francisco, San Francisco, California.
authors.forename: Pamela
authors.initials: P
authors.lastname: Yango
authors.affiliation: Department of Obstetrics, Gynecology and Reproductive Sciences, University of California, San Francisco, San Francisco, California; Helen Schneider Hospital for Women, Rabin Medical Center, Petah-Tiqva, Israel.
authors.forename: Eran
authors.initials: E
authors.lastname: Altman
authors.affiliation: Department of Obstetrics, Gynecology and Reproductive Sciences, University of California, San Francisco, San Francisco, California; Department of Urology, University of California, San Francisco, San Francisco, California.
authors.forename: James F
authors.initials: JF
authors.lastname: Smith
authors.affiliation: Department of Obstetrics and Gynecology, Albert Einstein University, Bronx, New York.
authors.forename: Peter C
authors.initials: PC
authors.lastname: Klatsky
authors.affiliation: Department of Obstetrics, Gynecology and Reproductive Sciences, University of California, San Francisco, San Francisco, California. Electronic address: trann@obgyn.ucsf.edu.
authors.forename: Nam D
authors.initials: ND
authors.lastname: Tran
mesh_terms: D000328:Adult; D053687:Adult Stem Cells / Q000166:cytology* / Q000502:physiology*; D061249:Batch Cell Culture Techniques / Q000379:methods; D002452:Cell Count; D002469:Cell Separation / Q000379:methods*; D002470:Cell Survival / Q000502:physiology; D002478:Cells, Cultured; D015925:Cryopreservation / Q000379:methods*; D006801:Humans; D008297:Male; D008875:Middle Aged; D013076:Sperm Count; D013091:Spermatogenesis / Q000502:physiology; D013094:Spermatozoa / Q000166:cytology* / Q000502:physiology*; D013737:Testis / Q000166:cytology*
keywords: Testicular cell cryopreservation; spermatogonial stem cells; testicular tissue cryopreservation