The aim of this study was to elucidate the molecular status of single human adult germ stem cells (haGSCs) and haGSC colonies, which spontaneously developed from the CD49f MACS and matrix- (collagen-/laminin+ binding-) selected fraction of enriched spermatogonia. Single-cell transcriptional profiling by Fluidigm BioMark system of a long-term cultured haGSCs cluster in comparison to human embryonic stem cells (hESCs) and human fibroblasts (hFibs) revealed that haGSCs showed a characteristic germ- and pluripotency-associated gene expression profile with some similarities to hESCs and with a significant distinction from somatic hFibs. Genome-wide comparisons with microarray analysis confirmed that different haGSC colonies exhibited gene expression heterogeneity with more or less pluripotency. The results of this study confirm that haGSCs are adult stem cells with a specific molecular gene expression profile in vitro, related but not identical to true pluripotent stem cells. Under ES-cell conditions haGSC colonies could be selected and maintained in a partial pluripotent state at the molecular level, which may be related to their cell plasticity and potential to differentiate into cells of all germ layers.

Department of Urology University of Tübingen Hospital Hoppe Seyler Straße 3 72076 Tübingen Germany

Institute for Anatomy and Cell Biology Medical Faculty University of Heidelberg Im Neuenheimer Feld 307 69120 Heidelberg Germany

Institute for Anatomy and Cell Biology Medical Faculty University of Heidelberg Im Neuenheimer Feld 307 69120 Heidelberg Germany ; Department of Stem Cells and Developmental Biology Cell Science Research Center Royan Institute for Stem Cell Biology and Technology P O Box 19395 Tehran 4644 Iran ; Faculty of Biotechnology Amol University of Special Modern Technologies P O Box 46168 Amol 49767 Iran

Institute of Anthropology and Human Genetics Microarray Facility University Clinic Calwerstraße 7 72076 Tübingen Germany

Sabine Conrad P O Box 12 43 72072 Tübingen Germany

TATAA Biocenter AB Odinsgatan 28 41103 Göteborg Sweden ; Institute of Biotechnology at the Czech Academy of Sciences Videnska 1083 14220 Prague 4 Czech Republic

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