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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

Immunofluorescence localization of STELLA and DAZL in human umbilical cord mesenchymal stem cell (HUMSC)-derived germ cell-like cells. Staining for STELLA (red) (A,B,E) and DAZL (red) (C, D, F) in differentiated cells (A, C, E, F) on day 14 after co-culture and HUMSCs cultured alone (B, D) were performed and examined by immunofluorescence microscopy (A-D) and confocal microscopy (E, F). Nuclei were counterstained with DAPI (blue). Arrowheads indicate nuclear staining, and arrows indicate cytoplasmic staining. (Magnification 200×).
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Fig5: Immunofluorescence localization of STELLA and DAZL in human umbilical cord mesenchymal stem cell (HUMSC)-derived germ cell-like cells. Staining for STELLA (red) (A,B,E) and DAZL (red) (C, D, F) in differentiated cells (A, C, E, F) on day 14 after co-culture and HUMSCs cultured alone (B, D) were performed and examined by immunofluorescence microscopy (A-D) and confocal microscopy (E, F). Nuclei were counterstained with DAPI (blue). Arrowheads indicate nuclear staining, and arrows indicate cytoplasmic staining. (Magnification 200×).

Mentions: We next examined the expression and localization of human-specific STELLA and DAZL proteins in clump-forming male germ cell-like cells after 14-day culture by immunofluorescence and confocal microscopy (Figure 5). We chose this time point to assess protein expression because the mRNAs for both germ cell markers were expressed at this stage. Both markers were detectable in co-cultured HUMSCs, but not in HUMSCs cultured alone (Figure 5B, D). Expression of human-specific STELLA in co-cultured HUMSCs was found to be primarily in cytoplasmic and nuclear regions, while the DAZL protein was localized to the nucleus.Figure 5


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)

Immunofluorescence localization of STELLA and DAZL in human umbilical cord mesenchymal stem cell (HUMSC)-derived germ cell-like cells. Staining for STELLA (red) (A,B,E) and DAZL (red) (C, D, F) in differentiated cells (A, C, E, F) on day 14 after co-culture and HUMSCs cultured alone (B, D) were performed and examined by immunofluorescence microscopy (A-D) and confocal microscopy (E, F). Nuclei were counterstained with DAPI (blue). Arrowheads indicate nuclear staining, and arrows indicate cytoplasmic staining. (Magnification 200×).
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Fig5: Immunofluorescence localization of STELLA and DAZL in human umbilical cord mesenchymal stem cell (HUMSC)-derived germ cell-like cells. Staining for STELLA (red) (A,B,E) and DAZL (red) (C, D, F) in differentiated cells (A, C, E, F) on day 14 after co-culture and HUMSCs cultured alone (B, D) were performed and examined by immunofluorescence microscopy (A-D) and confocal microscopy (E, F). Nuclei were counterstained with DAPI (blue). Arrowheads indicate nuclear staining, and arrows indicate cytoplasmic staining. (Magnification 200×).
Mentions: We next examined the expression and localization of human-specific STELLA and DAZL proteins in clump-forming male germ cell-like cells after 14-day culture by immunofluorescence and confocal microscopy (Figure 5). We chose this time point to assess protein expression because the mRNAs for both germ cell markers were expressed at this stage. Both markers were detectable in co-cultured HUMSCs, but not in HUMSCs cultured alone (Figure 5B, D). Expression of human-specific STELLA in co-cultured HUMSCs was found to be primarily in cytoplasmic and nuclear regions, while the DAZL protein was localized to the nucleus.Figure 5

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