Rat Sertoli cells express epithelial but also mesenchymal genes after immortalization with SV40

Konrad, Lutz; Keilani, Marcel Munir; Cordes, Andrea; Völck-Badouin, Elke; Laible, Leslie; Albrecht, Martín; Renneberg, Heiner; Aumüller, Gerhard

doi:10.1016/j.bbagen.2004.10.015

Cited by 24 publications

(23 citation statements)

References 47 publications

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“…Only Sertoli cell lines prepared from neonatal mice maintain AR expression (Mather 1980). AR expression is lost in immortalised rat Sertoli cell lines established through SV40-induced transformation (Konrad et al 2005) and in most Sertoli cell lines established from the cells of adult mouse testes via gene transfer. Therefore, transgenic techniques may not be adequate for maintaining AR expression in immortalised Sertoli cells.…”

Section: Discussionmentioning

confidence: 98%

“…Several cell lines have been established by the immortalisation of primary cultured mature Sertoli cells using the temperaturesensitive SV40 or the polyoma T-antigen transgenic mouse (Rassoulzadegan et al 1993, Tabuchi et al 2002. However, several problems have occurred during the establishment of cell lines after gene engineering; for example, after immortalisation, rat Sertoli cell lines expressed epithelial genes, but also unexpectedly expressed mesenchymal genes (Konrad et al 2005).…”

Section: Introductionmentioning

confidence: 99%

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Establishment of adult mouse Sertoli cell lines by using the starvation method

Sato¹,

Yoshida²,

Nozawa³

et al. 2013

Reproduction

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Sertoli cells were isolated from the testes of 6-week-old mice and stable Sertoli cell lines with higher proliferation rates were subcloned after starvation of primary cultured cells. After two rounds of this subcloning, 33 subcloned lines were selected on the basis of their proliferation rates. In addition, these subclones were screened according to their phagocytic activity and the characteristics of mature Sertoli cells, such as the expression of androgen receptors (ARs) and progesterone receptors, by using western blotting and immunocytochemical analysis, in addition to their morphology and proliferation rates. After the third round of subcloning, 12 subclones were selected for the final selection using RT-PCR for identification of genes specifically expressed by various testicular cells. Three clones were selected that expressed Sertoli-cell-specific genes, i.e. stem cell factor, clusterin, AR, a-inhibin, transferrin, Wilms' tumour-1, Mü llerian inhibitory substance, sex-determining region Y-box 9, FSH receptor (Fshr) and occludin; however, these clones did not express globulin transcription factor 1, steroidogenic factor or androgen-binding protein. These clones also expressed growth and differentiation factors that act on germ cells, such as leukaemia inhibitory factor, transforming growth factor b1 and basic fibroblast growth factor 2, but did not express c-kit (specific for germ cells), LH receptor and 3b-hydroxyl-dehydrogenase (specific for Leydig cells). Immunocytochemical data confirmed the expression of clusterin in these clones. Furthermore, the Bromodeoxyuridine incorporation assay confirmed the proliferation activity of these clones through Fshr after treatment with FSH. These clones are considered to be valuable tools for the study of Sertoli cell-specific gene expression and function. Reproduction (2013) 145 505-516

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Section: Discussionmentioning

confidence: 98%

Section: Introductionmentioning

confidence: 99%

Establishment of adult mouse Sertoli cell lines by using the starvation method

Sato¹,

Yoshida²,

Nozawa³

et al. 2013

Reproduction

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“…Similarly the rat Sertoli cell line SCIT-C8 expresses the epithelial markers transferrin, clusterin and stem cell factor but has lost androgen receptor expression and androgen responsiveness whereas it has aquired expression of some mesenchymal markers like fibronectin and entactin-1 33 …”

Section: Discussionmentioning

confidence: 99%

Isolation of Sertoli Cells and Peritubular Cells from Rat Testes

Bhushan

Aslani

Zhang

et al. 2016

JoVE

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The testis, and in particular the male gamete, challenges the immune system in a unique way because differentiated sperm first appear at the time of puberty -more than ten years after the establishment of systemic immune tolerance. Spermatogenic cells express a number of proteins that may be seen as non-self by the immune system. The testis must then be able to establish tolerance to these neo-antigens on the one hand but still be able to protect itself from infections and tumor development on the other hand. Therefore the testis is one of a few immune privileged sites in the body that tolerate foreign antigens without evoking a detrimental inflammatory immune response. Sertoli cells play a key role for the maintenance of this immune privileged environment of the testis and also prolong survival of cotransplanted cells in a foreign environment. Therefore primary Sertoli cells are an important tool for studying the immune privilege of the testis that cannot be easily replaced by established cell lines or other cellular models. Here we present a detailed and comprehensive protocol for the isolation of Sertoli cells -and peritubular cells if desired -from rat testes within a single day. Video LinkThe video component of this article can be found at

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“…Wagoner et al (2005) showed low levels of a4 isoform in 1-week-old rat testis using a preparation enriched in germ cell types by means of unit gravity sedimentation method, thus preventing the interference of other testis cells, and in spermatogonia isolated from 10-day-old rats. In contrast, Konrad et al (2011) detected the mRNA for Na C /K C -ATPase a1 and a4 isoforms in prepubertal rat-derived Sertoli cell line 93RS2 and in pubertal Sertoli cell line SCIT-C8, both immortalized cells (Jiang et al 1997, Konrad et al 2005. It is important to emphasize that, different from primary Sertoli cells, the immortalized cells fail to express androgen receptors and to respond to androgens and FSH (Konrad et al 2005).…”

Section: Ouabain Signaling In Rat Sertoli Cellsmentioning

confidence: 99%

“…In contrast, Konrad et al (2011) detected the mRNA for Na C /K C -ATPase a1 and a4 isoforms in prepubertal rat-derived Sertoli cell line 93RS2 and in pubertal Sertoli cell line SCIT-C8, both immortalized cells (Jiang et al 1997, Konrad et al 2005. It is important to emphasize that, different from primary Sertoli cells, the immortalized cells fail to express androgen receptors and to respond to androgens and FSH (Konrad et al 2005). The mRNA for Na C /K CATPase a4 isoform was also detected in primary culture of Sertoli cell from 19-day-old rat (Konrad et al 2011); at this age, rat Sertoli cells start differentiation.…”

Section: Ouabain Signaling In Rat Sertoli Cellsmentioning

confidence: 99%

Na+/K+-ATPase α1 isoform mediates ouabain-induced expression of cyclin D1 and proliferation of rat Sertoli cells

et al. 2012

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Novel roles for the interaction of cardiotonic steroids to Na H]thymidine, both of which were dependent on MAPK3/1 but not AKT intracellular cascade, as shown by pretreatment with MEK (MAP2K1/2) inhibitor U0126 and PI3K inhibitor wortmannin respectively. Moreover, the effect of ouabain on these proliferation parameters was completely prevented by phospho-cAMP response element-binding protein (CREB)/CREB-binding protein complex inhibitor KG501 and only partially by nuclear factor kB nuclear translocation inhibitor SN50. Pretreatment with estrogen receptor antagonist ICI 182 780 showed that MAPK3/1 activation by ouabain does not involve this receptor. The Na C /K C -ATPase a1 isoform, but not a4, was detected in Sertoli cells, suggesting that ouabain effects in Sertoli cells are mediated via a1. Taken together, these results show a rapid ouabain action in the Sertoli cells, which in turn can modulate nuclear transcriptional events essential for Sertoli cell proliferation in a critical period of testicular development. Our findings are important to understand the role of ouabain in the testis and its possible implications in male infertility. Reproduction (2012) 144 737-745

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Rat Sertoli cells express epithelial but also mesenchymal genes after immortalization with SV40

Cited by 24 publications

References 47 publications

Establishment of adult mouse Sertoli cell lines by using the starvation method

Establishment of adult mouse Sertoli cell lines by using the starvation method

Isolation of Sertoli Cells and Peritubular Cells from Rat Testes

Na+/K+-ATPase α1 isoform mediates ouabain-induced expression of cyclin D1 and proliferation of rat Sertoli cells

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