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Biomed Res Int. 2014;2014:138350. doi: 10.1155/2014/138350. Epub 2014 Mar 12.

Differential gene expression profiling of enriched human spermatogonia after short- and long-term culture.

Author information

  • 1Institute of Anatomy, University of Tübingen, Österbergstraße 3, 72074 Tübingen, Germany.
  • 2Institute for Anatomy and Cell Biology, Medical Faculty, University of Heidelberg, Im Neuenheimer Feld 307, 69120 Heidelberg, Germany ; Amol University of Special Modern Technologies, Special Modern Technologies, P.O. Box 46168-49767, Amol, Iran ; Department of Stem Cells and Developmental Biology, Royan Institute, P.O. Box 19395-4644, Tehran, Iran.
  • 3Institute for Anatomy and Cell Biology, Medical Faculty, University of Heidelberg, Im Neuenheimer Feld 307, 69120 Heidelberg, Germany.
  • 4TATAA Biocenter AB, Odinsgatan 28, 41103 Göteborg, Sweden and Institute of Biotechnology at the Czech Academy of Sciences, Vídenská 1083, 14220 Prague 4, Czech Republic.
  • 5Institute of Anthropology and Human Genetics, Microarray Facility, University Clinic, Calwerstraße 7, 72076 Tübingen, Germany.
  • 6Department of Urology, University Clinic Tübingen, Hoppe-Seyler-Straße 3, 72076 Tübingen, Germany.

Abstract

This study aimed to provide a molecular signature for enriched adult human stem/progenitor spermatogonia during short-term (<2 weeks) and long-term culture (up to more than 14 months) in comparison to human testicular fibroblasts and human embryonic stem cells. Human spermatogonia were isolated by CD49f magnetic activated cell sorting and collagen(-)/laminin(+) matrix binding from primary testis cultures obtained from ten adult men. For transcriptomic analysis, single spermatogonia-like cells were collected based on their morphology and dimensions using a micromanipulation system from the enriched germ cell cultures. Immunocytochemical, RT-PCR and microarray analyses revealed that the analyzed populations of cells were distinct at the molecular level. The germ- and pluripotency-associated genes and genes of differentiation/spermatogenesis pathway were highly expressed in enriched short-term cultured spermatogonia. After long-term culture, a proportion of cells retained and aggravated the "spermatogonial" gene expression profile with the expression of germ and pluripotency-associated genes, while in the majority of long-term cultured cells this molecular profile, typical for the differentiation pathway, was reduced and more genes related to the extracellular matrix production and attachment were expressed. The approach we provide here to study the molecular status of in vitro cultured spermatogonia may be important to optimize the culture conditions and to evaluate the germ cell plasticity in the future.

PMID:
24738045
[PubMed - in process]
PMCID:
PMC3971551
Free PMC Article

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