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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 human umbilical cord mesenchymal stem cells (HUMSCs) co-cultured with Sertoli cells (SCs). (A-E) HUMSC-derived germ cell-like cell colonies formed on a monolayer of SCs at specified time points after co-culture. (F) Morphology of SCs on day 28 after culture without HUMSCs. (Magnification 100×).
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Fig3: Morphology of human umbilical cord mesenchymal stem cells (HUMSCs) co-cultured with Sertoli cells (SCs). (A-E) HUMSC-derived germ cell-like cell colonies formed on a monolayer of SCs at specified time points after co-culture. (F) Morphology of SCs on day 28 after culture without HUMSCs. (Magnification 100×).

Mentions: Isolated HUMSCs were seeded on SC feeder layers. After 24-hour seeding, the transferred cells spread on top of the SC monolayer (Figure 3A). Morphological changes in HUMSCs were observed under a phase-contrast microscope every 3 days. Approximately 1 week later, HUMSCs lost their spindle-like shape, and a few small, loose colonies could be seen (Figure 3B). On day 14, more colonies with germ cell features appeared, and the size of clumped germ cell-like cells seemed larger (Figure 3C). The clumped germ cell-like cells were tightly packed and became flat on day 21 (Figure 3D). On day 28, the colonies showed a typically round shape (Figure 3E). No such changes were observed in untreated SCs or HUMSCs cultured alone (Figure 3F). At passage 5, HUMSCs cultured alone were maintained as spheres.Figure 3


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 human umbilical cord mesenchymal stem cells (HUMSCs) co-cultured with Sertoli cells (SCs). (A-E) HUMSC-derived germ cell-like cell colonies formed on a monolayer of SCs at specified time points after co-culture. (F) Morphology of SCs on day 28 after culture without HUMSCs. (Magnification 100×).
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Fig3: Morphology of human umbilical cord mesenchymal stem cells (HUMSCs) co-cultured with Sertoli cells (SCs). (A-E) HUMSC-derived germ cell-like cell colonies formed on a monolayer of SCs at specified time points after co-culture. (F) Morphology of SCs on day 28 after culture without HUMSCs. (Magnification 100×).
Mentions: Isolated HUMSCs were seeded on SC feeder layers. After 24-hour seeding, the transferred cells spread on top of the SC monolayer (Figure 3A). Morphological changes in HUMSCs were observed under a phase-contrast microscope every 3 days. Approximately 1 week later, HUMSCs lost their spindle-like shape, and a few small, loose colonies could be seen (Figure 3B). On day 14, more colonies with germ cell features appeared, and the size of clumped germ cell-like cells seemed larger (Figure 3C). The clumped germ cell-like cells were tightly packed and became flat on day 21 (Figure 3D). On day 28, the colonies showed a typically round shape (Figure 3E). No such changes were observed in untreated SCs or HUMSCs cultured alone (Figure 3F). At passage 5, HUMSCs cultured alone were maintained as spheres.Figure 3

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