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Journal of Biomedicine and Biotechnology
Volume 2012 (2012), Article ID 291038, 15 pages
Research Article

Potential Stemness of Frozen-Thawed Testicular Biopsies without Sperm in Infertile Men Included into the In Vitro Fertilization Programme

1Reproductive Unit, Department of Obstetrics and Gynaecology, University Medical Centre Ljubljana, 1000 Ljubljana, Slovenia
2Institute for Anatomy and Cell Biology, School of Medicine, University of Heidelberg, 69120 Heidelberg, Germany
3TATAA Biocenter, 41103 Göteborg, Sweden
4Laboratory of Gene Expression, Institute of Biotechnology, Academy of Sciences of the Czech Republic, 14220 Prague, Czech Republic
5Blood Transfusion Centre of Slovenia, 1000 Ljubljana, Slovenia
6Institute for Anatomy, School of Medicine, University of Tübingen, 72074 Tübingen, Germany

Received 31 May 2011; Revised 28 October 2011; Accepted 30 October 2011

Academic Editor: Ji Wu

Copyright © 2012 Martin Stimpfel et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.


We describe the potential stemness of a small amount of frozen-thawed testicular tissue without sperm obtained by biopsy from six patients undergoing assisted reproductive treatment. The patients were diagnosed with Sertoli Cell-Only Syndrome alone or combined with maturation arrest. Trying to provide the natural stem cell niche for cultured stem cells, all isolated cells from enzymatically degraded biopsies where cultured together in different culture media and the presence of putative mesenchymal and putative pluripotent ES-like stem cells was indicated using different methods. High throughput real-time quantitative PCR followed by multivariate analysis revealed the formation of distinct cell clusters reflecting high degree of similarity and some of these cell clusters expressed the genes characteristic for pluripotent stem cells. In the presence of the follicular fluid, prepared as serum, putative testicular stem cells showed a certain degree of plasticity, and spontaneously differentiated into adipose-like and neuronal-like cells. Additionally, using differentiation protocols putative testicular stem cells were differentiated into neuronal- and pancreatic-like cells. This study shows that in assisted reproduction programmes, testicular tissue with no sperm might be an important source of stem cells, although it is discarded in daily medical practice; this requires further research.