Reassembly of adult human testicular cells: can testis cord-like structures be created in vitro?

Mincheva, M; Sandhowe-Klaverkamp, Reinhild; Wistuba, Joachim; Redmαnn, K.; Stukenborg, Jan-Bernd; Kliesch, Sabine; Schlatt, Stefan

doi:10.1093/molehr/gax063

Cited by 27 publications

(9 citation statements)

References 42 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…These genes, including KRT18 , PTGDS and SOX9 , are used as markers of Sertoli cells or are highly expressed in Sertoli cells (Buganim et al, 2012; Sharpe et al, 2003). We further compared the expression of 59 genes that are highly expressed in Sertoli cells (Bouma et al, 2010; Boyer et al, 2004; Mincheva et al, 2018) among the 2F-hiSCs, 5F-hiSCs and aSCs. All three groups exhibited similar expression of many markers that are expressed in more mature Sertoli cells, including CDKN1B (or p27 kip1 ) and CLU (Wang et al, 2016).…”

Section: Resultsmentioning

confidence: 99%

Induction of Sertoli-like cells from human fibroblasts by NR5A1 and GATA4

Liang

Wang

et al. 2019

eLife

View full text Add to dashboard Cite

Sertoli cells are essential nurse cells in the testis that regulate the process of spermatogenesis and establish the immune-privileged environment of the blood-testis-barrier (BTB). Here, we report the in vitro reprogramming of fibroblasts to human induced Sertoli-like cells (hiSCs). Initially, five transcriptional factors and a gene reporter carrying the AMH promoter were utilized to obtain the hiSCs. We further reduce the number of reprogramming factors to two, NR5A1 and GATA4, and show that these hiSCs have transcriptome profiles and cellular properties that are similar to those of primary human Sertoli cells. Moreover, hiSCs can sustain the viability of spermatogonia cells harvested from mouse seminiferous tubules. hiSCs suppress the proliferation of human T lymphocytes and protect xenotransplanted human cells in mice with normal immune systems. hiSCs also allow us to determine a gene associated with Sertoli cell only syndrome (SCO), CX43, is indeed important in regulating the maturation of Sertoli cells.

show abstract

Section: Resultsmentioning

confidence: 99%

Induction of Sertoli-like cells from human fibroblasts by NR5A1 and GATA4

Liang

Wang

et al. 2019

eLife

View full text Add to dashboard Cite

Sertoli cells are essential nurse cells in the testis that regulate the process of spermatogenesis and establish the immune-privileged environment of the blood-testis-barrier (BTB). Here, we report the in vitro reprogramming of fibroblasts to human induced Sertoli-like cells (hiSCs). Initially, five transcriptional factors and a gene reporter carrying the AMH promoter were utilized to obtain the hiSCs. We further reduce the number of reprogramming factors to two, NR5A1 and GATA4, and show that these hiSCs have transcriptome profiles and cellular properties that are similar to those of primary human Sertoli cells. Moreover, hiSCs can sustain the viability of spermatogonia cells harvested from mouse seminiferous tubules. hiSCs suppress the proliferation of human T lymphocytes and protect xenotransplanted human cells in mice with normal immune systems. hiSCs also allow us to determine a gene associated with Sertoli cell only syndrome (SCO), CX43, is indeed important in regulating the maturation of Sertoli cells.

show abstract

“…However, the use of engineering methods to generate de novo testicular tissues entirely in vitro has been comparably more challenging. In addition, to our knowledge only three published testicular organoid models have explored using adult primary cells for organoid generation; the greater majority of reports use either immortalized, pre-pubertal, or otherwise immature cells [15,57,58]. Expanding organoid generation protocols to include adult cells, especially human adult cells, is a necessary step for applying organoid systems towards the study of human reproductive health, infertility, and disease.…”

Section: Discussionmentioning

confidence: 99%

Testicular organoid formation is a property of immature somatic cells, which self-assemble and exhibit long-term hormone-responsive endocrine function

Edmonds

Woodruff

2020

Biofabrication

View full text Add to dashboard Cite

Testicular organoid models are tools to study testicular physiology, development, and spermatogenesis in vitro. However, few side-by-side comparisons of organoid generation method have been evaluated. Here, we directly tested whether the culture microenvironment is the prime determinant promoting testicular organoid self-assembly. Using Matrigel as a representative extracellular matrix (ECM), we compared multiple culture environments, 2D and 3D, ECM-free and ECM, for organoid self-assembly with immature murine testicular cells. De novo tissues were observed to self-assemble in all four culture environments tested within 72 h, however, these tissues only met requirements to be named organoids in 2D ECM and 3D ECM-free (3DF) culture methods. Based on these results, 3DF was selected for further study, and used to examine animal age as an independent variable. Organoid assembly was significantly delayed when using pubertal murine cells and entirely absent from adult murine and adult human cells. Organoid-conditioned medium and medium supplemented with 1% Matrigel did not improve organoid assembly in pubertal murine cells, but immature murine cells rescued the assembly of adult murine cells when cultured together as age-chimeric cell mixtures. In murine organoids cultured for 14 d, tubule-like structures exhibiting a highly biomimetic architecture were characterized, including some rare germ and spermatogonial stem cells. These structural organoids secreted high levels of testosterone and inhibin B over 12 weeks with preserved responsivity to gonadotropins. Collectively these studies, in which cellular self-assembly and organoid formation was achieved independent of the culture microenvironment, suggest that self-assembly is an innate property of immature testicular cells independent from, but capable of being promoted by, the culture environment. This study provides a template for studying testicular organoid self-assembly and endocrine function, and a platform for improving the engineering of functional testicular tissues.

show abstract

“…These genes, including KRT18, PTGDS and SOX9 , are used as markers of Sertoli cells or are highly expressed in Sertoli cells 26,35 . We further compared the expression of 59 genes that are highly expressed in Sertoli cells 36–38 among the 2F-hiSCs, 5F-hiSCs and aSCs. All three groups exhibited similar expression of many markers that are expressed in more mature Sertoli cells, including CDKN1B (or p27 kip1 ) and CLU 39 .…”

Section: Resultsmentioning

confidence: 99%

Induction of Sertoli cells from human fibroblasts by NR5A1 and GATA4

Liang

Wang

et al. 2018

Preprint

View full text Add to dashboard Cite

Sertoli cells are essential nurse cells in the testis that regulate the process of spermatogenesis and establish the immune-privileged environment of the blood-testis-barrier (BTB). The induction of human Sertoli cells from fibroblasts could provide cellular sources for fertility and transplantation treatments. Here, we report the in vitro reprogramming of human fibroblasts to Sertoli cells and characterize these human induced Sertoli cells (hiSCs). Initially, five transcriptional factors (NR5A1, GATA4, WT1, SOX9 and DMRT1) and a gene reporter carrying the AMH promoter were utilized to obtain the hiSCs. We further reduce the number of reprogramming factors to two, i.e., NR5A1 and GATA4, and show that these hiSCs have transcriptome profiles that are similar to those of primary human Sertoli cells. Consistent with the known cellular properties of Sertoli cells, hiSCs attract endothelial cells and exhibit high number of lipid droplets in the cytoplasm. More importantly, hiSCs can sustain the viability of spermatogonia cells harvested from mouse seminiferous tubules. In addition, hiSCs suppress the production of IL-2 and proliferation of human T lymphocytes. When hiSCs were cotransplanted with human embryonic kidney cells, these xenotransplanted human cells survived longer in mice with normal immune systems. hiSCs also allow us to determine a gene associated with Sertoli-only syndrome (SCO), CX43, is indeed important in regulating the maturation of Sertoli cells.

show abstract

Reassembly of adult human testicular cells: can testis cord-like structures be created in vitro?

Abstract: The work was supported by EU-FP7-PEOPLE-2013-ITN 603568: 'Growsperm'. No conflict of interests is declared.

Cited by 27 publications

References 42 publications

Induction of Sertoli-like cells from human fibroblasts by NR5A1 and GATA4

Induction of Sertoli-like cells from human fibroblasts by NR5A1 and GATA4

Testicular organoid formation is a property of immature somatic cells, which self-assemble and exhibit long-term hormone-responsive endocrine function

Induction of Sertoli cells from human fibroblasts by NR5A1 and GATA4

Contact Info

Product

Resources

About