title: Enrichment of spermatogonial stem cells from long-term cultured human testicular cells.
abstract: OBJECTIVE
  To evaluate the degree of enrichment of spermatogonial stem cells (SSCs) from human testicular cell cultures by ITGA6+, HLA-/ITGA6+, GPR125+, and HLA-/GPR125+ magnetic-assisted cell sorting (MACS).
  
  
  METHODS
  Experimental basic science study.
  
  
  METHODS
  Reproductive biology laboratory.
  
  
  METHODS
  Multiple samples of cryopreserved human testicular cells from two prostate cancer patients with normal spermatogenesis.
  
  
  METHODS
  Cultured human testicular cells subjected to four sorting strategies based on MACS and xenotransplanted to the testes of mice to determine the enrichment for SSCs.
  
  
  METHODS
  Enrichment for human spermatogonia and SSCs tested by expression analysis of spermatogonial markers ITGA6, GPR125, ZBTB16, UCHL1, and ID4 using quantitative real-time polymerase chain reaction (qPCR) and by xenotransplantation into the testes of mice, respectively.
  
  
  RESULTS
  Compared with the nonsorted cultured testicular cells, only the ITGA6+ and HLA-/GPR125+ sorted cells showed enrichment for ID4. No difference in expression of ZBTB16 and UCHL1 was observed. Xenotransplantation of the sorted cell fractions showed a 7.1-fold enrichment of SSCs with ITGA6+.
  
  
  CONCLUSIONS
  Magnetic-assisted cell sorting of cultured human testicular cells using ITGA6 allows for enrichment of SSCs, which aids in further molecular characterization of cultured human SSCs and enhances testicular colonization upon transplantation in future clinical settings.
authors.affiliation: Center for Reproductive Medicine, Women's and Children's Hospital, Academic Medical Center, University of Amsterdam, Amsterdam, the Netherlands.
authors.forename: Bita
authors.initials: B
authors.lastname: Nickkholgh
authors.affiliation: Center for Reproductive Medicine, Women's and Children's Hospital, Academic Medical Center, University of Amsterdam, Amsterdam, the Netherlands.
authors.forename: Sefika Canan
authors.initials: SC
authors.lastname: Mizrak
authors.affiliation: Center for Reproductive Medicine, Women's and Children's Hospital, Academic Medical Center, University of Amsterdam, Amsterdam, the Netherlands.
authors.forename: Cindy M
authors.initials: CM
authors.lastname: Korver
authors.affiliation: Center for Reproductive Medicine, Women's and Children's Hospital, Academic Medical Center, University of Amsterdam, Amsterdam, the Netherlands.
authors.forename: Saskia K M
authors.initials: SK
authors.lastname: van Daalen
authors.affiliation: Center for Reproductive Medicine, Women's and Children's Hospital, Academic Medical Center, University of Amsterdam, Amsterdam, the Netherlands.
authors.forename: Andreas
authors.initials: A
authors.lastname: Meissner
authors.affiliation: Center for Reproductive Medicine, Women's and Children's Hospital, Academic Medical Center, University of Amsterdam, Amsterdam, the Netherlands.
authors.forename: Sjoerd
authors.initials: S
authors.lastname: Repping
authors.affiliation: Center for Reproductive Medicine, Women's and Children's Hospital, Academic Medical Center, University of Amsterdam, Amsterdam, the Netherlands. Electronic address: a.m.vanpelt@amc.uva.nl.
authors.forename: Ans M M
authors.initials: AM
authors.lastname: van Pelt
mesh_terms: D053687:Adult Stem Cells / Q000378:metabolism* / Q000637:transplantation; D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D002454:Cell Differentiation; D049109:Cell Proliferation; D002478:Cells, Cultured; D006680:HLA Antigens / Q000378:metabolism; D006801:Humans; D018189:Immunomagnetic Separation / Q000379:methods*; D051796:Inhibitor of Differentiation Proteins / Q000235:genetics / Q000378:metabolism; D039503:Integrin alpha6 / Q000235:genetics / Q000378:metabolism; D051741:Kruppel-Like Transcription Factors / Q000235:genetics / Q000378:metabolism; D008297:Male; D051379:Mice; D008819:Mice, Nude; D000075722:Promyelocytic Leukemia Zinc Finger Protein; D060888:Real-Time Polymerase Chain Reaction; D043562:Receptors, G-Protein-Coupled / Q000235:genetics / Q000378:metabolism; D013093:Spermatogonia / Q000378:metabolism* / Q000637:transplantation; D013737:Testis / Q000378:metabolism* / Q000637:transplantation; D013997:Time Factors; D014183:Transplantation, Heterologous; D043222:Ubiquitin Thiolesterase / Q000235:genetics / Q000378:metabolism
chemical_list: C548160:ADGRA3 protein, human; D015415:Biomarkers; D006680:HLA Antigens; C490128:ID4 protein, human; C588002:ITGA6 protein, human; D051796:Inhibitor of Differentiation Proteins; D039503:Integrin alpha6; D051741:Kruppel-Like Transcription Factors; D000075722:Promyelocytic Leukemia Zinc Finger Protein; D043562:Receptors, G-Protein-Coupled; C482860:UCHL1 protein, human; C092037:ZBTB16 protein, human; D043222:Ubiquitin Thiolesterase
keywords: Autotransplantation; ITGA6; enrichment; spermatogonial stem cells