In vitro generation of Sertoli-like and haploid spermatid-like cells from human umbilical cord perivascular cells
Autor: | Clifford Librach, Tanya Barretto, Ekaterina Shlush, Shlomit Kenigsberg, Sonja A. Swanson, Andrée Gauthier-Fisher, Leila Maghen, Sergey I. Moskovtsev |
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Jazyk: | angličtina |
Předmět: |
Male
0301 basic medicine Cellular differentiation Primary Cell Culture Gene Expression Medicine (miscellaneous) Mice SCID Human mesenchymal stem cell Biology Biochemistry Genetics and Molecular Biology (miscellaneous) Mice 03 medical and health sciences 0302 clinical medicine Paracrine Communication Niche medicine Animals Humans Spermatogenesis Cell potency Sertoli Cells Gene Expression Profiling Research Xenograft Ex-vivo spermatogenesis Cell Differentiation Mesenchymal Stem Cells Amniotic stem cells Cell Biology Fetal Blood Human umbilical cord Spermatids Sertoli cell Embryonic stem cell Cell biology 030104 developmental biology medicine.anatomical_structure Paracrine 030220 oncology & carcinogenesis Amniotic epithelial cells Heterografts Molecular Medicine Perivascular cell Stem cell Spermatogonial stem cell Germ cell Biomarkers Adult stem cell |
Zdroj: | Stem Cell Research & Therapy |
ISSN: | 1757-6512 |
DOI: | 10.1186/s13287-017-0491-8 |
Popis: | Background First trimester (FTM) and term human umbilical cord-derived perivascular cells (HUCPVCs), which are rich sources of mesenchymal stem cells (MSCs), can give rise to Sertoli cell (SC)-like as well as haploid germ cell (GC)-like cells in vitro using culture conditions that recapitulate the testicular niche. Gamete-like cells have been produced ex vivo using pluripotent stem cells as well as MSCs. However, the production of functional gametes from human stem cells has yet to be achieved. Methods Three independent lines of FTM and term HUCPVCs were cultured using a novel 5-week step-wise in vitro differentiation protocol recapitulating key physiological signals involved in testicular development. SC- and GC-associated phenotypical properties were assessed by real-time polymerase chain reaction (RT-PCR), quantitative PCR immunocytochemistry, flow cytometry, and fluorescence in-situ hybridization (FISH). Functional spermatogonial stem cell-like properties were assessed using a xenotranplantation assay. Results Within 3 weeks of differentiation, two morphologically distinct cell types emerged including large adherent cells and semi-attached round cells. Both early GC-associated markers (VASA, DAZL, GPR125, GFR1α) and SC-associated markers (FSHR, SOX9, AMH) were upregulated, and 5.7 ± 1.2% of these cells engrafted near the inner basal membrane in a xenograft assay. After 5 weeks in culture, 10–30% of the cells were haploid, had adopted a spermatid-like morphology, and expressed PRM1, Acrosin, and ODF2. Undifferentiated HUCPVCs secreted key factors known to regulate spermatogenesis (LIF, GDNF, BMP4, bFGF) and 10–20% of HUCPVCs co-expressed SSEA4, CD9, CD90, and CD49f. We hypothesize that the paracrine properties and cellular heterogeneity of HUCPVCs may explain their dual capacity to differentiate to both SC- and GC-like cells. Conclusions HUCPVCs recapitulate elements of the testicular niche including their ability to differentiate into cells with Sertoli-like and haploid spermatid-like properties in vitro. Our study supports the importance of generating a niche-like environment under ex vivo conditions aiming at creating mature GC, and highlights the plasticity of HUCPVCs. This could have future applications for the treatment of some cases of male infertility. Electronic supplementary material The online version of this article (doi:10.1186/s13287-017-0491-8) contains supplementary material, which is available to authorized users. |
Databáze: | OpenAIRE |
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