Establishment and applications of male germ cell and Sertoli cell lines

Wang, Hong; Wen, Lei; Yuan, Quan; Sun, Min; Niu, Minghui; He, Zuping

doi:10.1530/rep-15-0546

Cited by 38 publications

(25 citation statements)

References 68 publications

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“…In addition, Sertoli cells offer nutritional and immunological support for male germ cells26. Abnormal number and/or quality of Sertoli cells can lead to dyszoospermia27.…”

Section: Discussionmentioning

confidence: 99%

BMP6 Regulates Proliferation and Apoptosis of Human Sertoli Cells Via Smad2/3 and Cyclin D1 Pathway and DACH1 and TFAP2A Activation

Wang

Yuan

Sun

et al. 2017

Sci Rep

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Sertoli cells are essential for regulating normal spermatogenesis. However, the mechanisms underlying human Sertoli cell development remain largely elusive. Here we examined the function and signaling pathways of BMP6 in regulating human Sertoli cells. RT-PCR, immunocytochemistry and Western blots revealed that BMP6 and its multiple receptors were expressed in human Sertoli cells. CCK-8 and EDU assays showed that BMP6 promoted the proliferation of Sertoli cells. Conversely, BMP6 siRNAs inhibited the division of these cells. Annexin V/PI assay indicated that BMP6 reduced the apoptosis in human Sertoli cells, whereas BMP6 knockdown assumed reverse effects. BMP6 enhanced the expression levels of ZO1, SCF, GDNF and AR in human Sertoli cells, and ELISA assay showed an increase of SCF by BMP6 and a reduction by BMP6 siRNAs. Notably, Smad2/3 phosphorylation and cyclin D1 were enhanced by BMP6 and decreased by BMP6 siRNAs in human Sertoli cells. The levels of DACH1 and TFAP2A were increased by BMP6 and reduced by BMP6 siRNAs, and the growth of human Sertoli cells was inhibited by these siRNAs. Collectively, these results suggest that BMP6 regulates the proliferation and apoptosis of human Sertoli cells via activating the Smad2/3/cyclin D1 and DACH1 and TFAP2A pathway.

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“…In addition, Sertoli cells offer nutritional and immunological support for male germ cells26. Abnormal number and/or quality of Sertoli cells can lead to dyszoospermia27.…”

Section: Discussionmentioning

confidence: 99%

BMP6 Regulates Proliferation and Apoptosis of Human Sertoli Cells Via Smad2/3 and Cyclin D1 Pathway and DACH1 and TFAP2A Activation

Wang

Yuan

Sun

et al. 2017

Sci Rep

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“…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 . However, some markers in the aSCs, including NCAM2, INHA and KRT18 39,40 , were expressed at lower levels (Supplementary Fig.…”

Section: Resultsmentioning

confidence: 92%

“…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 . However, some markers in the aSCs, including NCAM2, INHA and KRT18 39,40 , were expressed at lower levels (Supplementary Fig. 3) than those in the other two hiSCs.…”

Section: Resultsmentioning

confidence: 92%

“…Ten marker genes previously reported to be more highly expressed in mature or immature Sertoli cells 35,39 were chosen for the expression level comparisons between WT and CX43KO hiSCs. The CX43KO hiSCs exhibited a higher expression of some immature markers and lower expression of mature markers, suggesting that the CX43KO cells were more immature than the WT hiSCs.…”

Section: Resultsmentioning

confidence: 99%

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Induction of Sertoli cells from human fibroblasts by NR5A1 and GATA4

Liang

Wang

et al. 2018

Preprint

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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.

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“…Testis Sertoli cell is the critical somatic cell lineages mediated SSC fate in vivo . Although most of the Sertoli lines established either by transgenic animals or gene infection had been widely applied for spermatogenesis mediation or investigation of Sertoli cells themselves272829, the application of Sertoli cells for SSC growth has not been validated. Recently testosterone-regulated GDNF secretion by peritubular myoid cells has also been observed, which ensured the optimal GDNF level for repopulating the undifferentiated spermatogonia including SSCs30.…”

Section: Discussionmentioning

confidence: 99%

GDNF-expressing STO feeder layer supports the long-term propagation of undifferentiated mouse spermatogonia with stem cell properties

Wei

Jia

Xue

et al. 2016

Sci Rep

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The development of a stem cell culture system would expedite our understanding of the biology of tissue regeneration. Spermatogonial stem cell (SSC) is the foundation for lifelong male spermatogenesis and the SSC culture has been optimized continuously in recent years. However, there have been many inconveniences to reconstruct SSC self-renewal and proliferation in vitro, such as the frequent refreshment of recombinant cytokines, including GDNF, the essential growth factor for SSC maintenance. In the present study, we observed that both STO and MEF cells, which were previously used as feeders for SSC growth, did not express GDNF, but a GDNF-expressing STO feeder could support undifferentiated mouse spermatogonia propagation in vitro for three months without the refreshment of recombinant growth factor GDNF. The cell morphology, growth rate and SSC-associated gene expression remained identical to the SSCs cultured using previous methods. The transplantation of SSCs growing on these GDNF-expressing STO feeders could generate extensive colonies of spermatogenesis in recipient testes, functionally validating the stemness of these cells. Collectively, our data indicated that the further modification of feeder cells might facilitate the self-renewal and propagation of SSCs in vitro.

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Establishment and applications of male germ cell and Sertoli cell lines

Cited by 38 publications

References 68 publications

BMP6 Regulates Proliferation and Apoptosis of Human Sertoli Cells Via Smad2/3 and Cyclin D1 Pathway and DACH1 and TFAP2A Activation

BMP6 Regulates Proliferation and Apoptosis of Human Sertoli Cells Via Smad2/3 and Cyclin D1 Pathway and DACH1 and TFAP2A Activation

Induction of Sertoli cells from human fibroblasts by NR5A1 and GATA4

GDNF-expressing STO feeder layer supports the long-term propagation of undifferentiated mouse spermatogonia with stem cell properties

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