Screening for biomarkers of spermatogonia within the human testis: a whole genome approach.
BACKGROUND
A key step in studying the biology of spermatogonia is to determine their global gene expression profile. However, disassociation of these cells from the testis may alter their profile to a considerable degree. To characterize the molecular phenotype of human spermatogonia, including spermatogonial stem cells (SSCs), within their cognate microenvironment, a rare subtype of human defective spermatogenesis was exploited in which spermatogonia were the only germ cell type.


METHODS
The global expression profile of these samples was assessed on the Affymetrix microarray platform and compared with tissues showing homogeneous Sertoli-cell-only appearance; selected genes were validated by quantitative real-time PCR and immunohistochemistry on disparate sample sets.


RESULTS
Highly significant differences in gene expression levels correlated with the appearance of spermatogonia, including 239 best candidates of human spermatogonially expressed genes. Specifically, fibroblast growth factor receptor 3 (FGFR3), desmoglein 2 (DSG2), E3 ubiquitin ligase c-CBL (casitas B-cell lymphoma), cancer/testis antigen NY-ESO-1 (CTAG1A/B), undifferentiated embryonic cell transcription factor 1 (UTF1) and synaptosomal-associated protein, 91 kDa homolog (SNAP91) were shown to represent specific biomarkers of human spermatogonia.


CONCLUSIONS
These biomarkers, specifically the surface markers FGFR3 and DSG2, may facilitate the isolation and enrichment of human stem and/or progenitor spermatogonia and thus lay a foundation for studies of long-term maintenance of human SSCs/progenitor cells, spermatogonial self-renewal, clonal expansion and differentiation.
von Kopylow
Kathrein
K
Department of Andrology, University Hospital Hamburg-Eppendorf, Hamburg, Germany.
Kirchhoff
Christiane
C
Jezek
Davor
D
Schulze
Wolfgang
W
Feig
Caroline
C
Primig
Michael
M
Steinkraus
Volker
V
Spiess
Andrej-Nikolai
AN
D053687:Adult Stem Cells / Q000166:cytology / Q000378:metabolism; D015415:Biomarkers / Q000378:metabolism; D002454:Cell Differentiation; D051301:Desmoglein 2 / Q000235:genetics / Q000378:metabolism; D020869:Gene Expression Profiling; D055106:Genome-Wide Association Study; D006801:Humans; D007150:Immunohistochemistry; D007248:Infertility, Male / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D008297:Male; D020411:Oligonucleotide Array Sequence Analysis; D025941:Protein Interaction Mapping; D051498:Receptor, Fibroblast Growth Factor, Type 3 / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013093:Spermatogonia / Q000166:cytology* / Q000378:metabolism*; D013737:Testis / Q000166:cytology* / Q000196:embryology / Q000378:metabolism*
D015415:Biomarkers; C495314:DSG2 protein, human; D051301:Desmoglein 2; C496365:FGFR3 protein, human; D051498:Receptor, Fibroblast Growth Factor, Type 3