Abstract
BackgroundMicroenvironment 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.MethodsOur 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.ResultsHUMSCs 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.ConclusionThe 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.
Highlights
Microenvironment signals play a critical role in directing the differentiation of stem cells
Morphology of human umbilical cord mesenchymal stem cells cultured alone Cultured HUMSCs appeared as spindle-shape cells migrating out from Wharton’s jelly fragments on day 5 to day 7 (Figure 1A)
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]
Summary
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. Considerable knowledge has been gained regarding germ cell development in the mouse, and many mechanisms are highly conserved in mammals, there remains a need to investigate human germ cell development directly [2]. In vitro culture systems of human germ cells. The research on derivation of male germ cells from stem cells has opened new perspectives for investigating germ cell development in vitro. The progression through the meiotic process is still a challenge in the in vitro differentiation of male germ cells from stem cells. The transfection of embryonic stem cell lines with marked or fluorescent proteins allows for characterization of the differentiated germ cells, but the use of transfected lines disqualifies the male germ cells obtained for their application in clinical procedures [6].
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