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Sertoli cell-mediated differentiation of male germ cell-like cells from human umbilical cord Wharton's jelly-derived mesenchymal stem cells in an in vitro co-culture system.

Xie L, Lin L, Tang Q, Li W, Huang T, Huo X, Liu X, Jiang J, He G, Ma L - Eur. J. Med. Res. (2015)

Bottom Line: Differentiated cells formed round colonies that share the morphological features of spermatogonial colonies.RT-PCR, immunofluorescence, confocal microscopy, and Western blot analyses revealed the expression of early germ cell markers STELLA and VASA and male germ cell-specific marker DAZL in differentiated HUMSCs, confirming the presence of cells with characteristics of male germ cells.Male germ cells derived from HUMSCs may be used in the therapy for male infertility.

View Article: PubMed Central - PubMed

Affiliation: Women's and Children's Hospital of Shenzhen University, Shenzhen, 518000, China. shirleyxie_2008@yahoo.cn.

ABSTRACT

Background: Microenvironment signals play a critical role in directing the differentiation of stem cells. Sertoli cells (SCs) provide a unique microenvironment that is essential for germ cell differentiation.

Methods: Our previous study has demonstrated that human umbilical cord Wharton's jelly-derived mesenchymal stem cells (HUMSCs) could differentiate towards male germ cells in vitro, but HUMSC-derived germ-like cells expressed only few germ cell markers. The aim of this study was to investigate the effect of SCs on the differentiation of HUMSCs towards male germ cells using a co-culture system that mimicked the in vivo male germ cell microenvironment.

Results: HUMSCs formed clump-like features on SC monolayers after seeding for 3 weeks. Differentiated cells formed round colonies that share the morphological features of spermatogonial colonies. RT-PCR, immunofluorescence, confocal microscopy, and Western blot analyses revealed the expression of early germ cell markers STELLA and VASA and male germ cell-specific marker DAZL in differentiated HUMSCs, confirming the presence of cells with characteristics of male germ cells.

Conclusion: The HUMSC-SC co-culture system mimics a native microenvironment for germ cell colonization without any in vitro artificial manipulation and can be used to explore the mechanisms controlling the differentiation of male germ cells from HUMSCs. Male germ cells derived from HUMSCs may be used in the therapy for male infertility.

No MeSH data available.


Related in: MedlinePlus

Morphology of cultured human umbilical cord mesenchymal stem cells (HUMSCs). (A) Primary HUMSCs on day 7 after culture. HUMSCs (black arrow) migrated out from Wharton’s jelly fragments (white arrow). (B) Fibroblast-like HUMSCs at passage 3. (Magnification 100×).
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Fig1: Morphology of cultured human umbilical cord mesenchymal stem cells (HUMSCs). (A) Primary HUMSCs on day 7 after culture. HUMSCs (black arrow) migrated out from Wharton’s jelly fragments (white arrow). (B) Fibroblast-like HUMSCs at passage 3. (Magnification 100×).

Mentions: Cultured HUMSCs appeared as spindle-shape cells migrating out from Wharton’s jelly fragments on day 5 to day 7 (Figure 1A). After passage, they appeared as fibroblast-like adherent cells and most were flat, wide and polygonal (Figure 1B). Assessment of the expression of markers associated with endothelial stem cells and adult stem cells by flow cytometry indicated that they possessed the multipotent characteristics of HUMSCs [23].Figure 1


Sertoli cell-mediated differentiation of male germ cell-like cells from human umbilical cord Wharton's jelly-derived mesenchymal stem cells in an in vitro co-culture system.

Xie L, Lin L, Tang Q, Li W, Huang T, Huo X, Liu X, Jiang J, He G, Ma L - Eur. J. Med. Res. (2015)

Morphology of cultured human umbilical cord mesenchymal stem cells (HUMSCs). (A) Primary HUMSCs on day 7 after culture. HUMSCs (black arrow) migrated out from Wharton’s jelly fragments (white arrow). (B) Fibroblast-like HUMSCs at passage 3. (Magnification 100×).
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4389972&req=5

Fig1: Morphology of cultured human umbilical cord mesenchymal stem cells (HUMSCs). (A) Primary HUMSCs on day 7 after culture. HUMSCs (black arrow) migrated out from Wharton’s jelly fragments (white arrow). (B) Fibroblast-like HUMSCs at passage 3. (Magnification 100×).
Mentions: Cultured HUMSCs appeared as spindle-shape cells migrating out from Wharton’s jelly fragments on day 5 to day 7 (Figure 1A). After passage, they appeared as fibroblast-like adherent cells and most were flat, wide and polygonal (Figure 1B). Assessment of the expression of markers associated with endothelial stem cells and adult stem cells by flow cytometry indicated that they possessed the multipotent characteristics of HUMSCs [23].Figure 1

Bottom Line: Differentiated cells formed round colonies that share the morphological features of spermatogonial colonies.RT-PCR, immunofluorescence, confocal microscopy, and Western blot analyses revealed the expression of early germ cell markers STELLA and VASA and male germ cell-specific marker DAZL in differentiated HUMSCs, confirming the presence of cells with characteristics of male germ cells.Male germ cells derived from HUMSCs may be used in the therapy for male infertility.

View Article: PubMed Central - PubMed

Affiliation: Women's and Children's Hospital of Shenzhen University, Shenzhen, 518000, China. shirleyxie_2008@yahoo.cn.

ABSTRACT

Background: Microenvironment signals play a critical role in directing the differentiation of stem cells. Sertoli cells (SCs) provide a unique microenvironment that is essential for germ cell differentiation.

Methods: Our previous study has demonstrated that human umbilical cord Wharton's jelly-derived mesenchymal stem cells (HUMSCs) could differentiate towards male germ cells in vitro, but HUMSC-derived germ-like cells expressed only few germ cell markers. The aim of this study was to investigate the effect of SCs on the differentiation of HUMSCs towards male germ cells using a co-culture system that mimicked the in vivo male germ cell microenvironment.

Results: HUMSCs formed clump-like features on SC monolayers after seeding for 3 weeks. Differentiated cells formed round colonies that share the morphological features of spermatogonial colonies. RT-PCR, immunofluorescence, confocal microscopy, and Western blot analyses revealed the expression of early germ cell markers STELLA and VASA and male germ cell-specific marker DAZL in differentiated HUMSCs, confirming the presence of cells with characteristics of male germ cells.

Conclusion: The HUMSC-SC co-culture system mimics a native microenvironment for germ cell colonization without any in vitro artificial manipulation and can be used to explore the mechanisms controlling the differentiation of male germ cells from HUMSCs. Male germ cells derived from HUMSCs may be used in the therapy for male infertility.

No MeSH data available.


Related in: MedlinePlus