Differentiation of spermatogonial stem cell-like cells from murine testicular tissue into haploid male germ cells in vitro

Wang, Peng; Suo, Li-Juan; Shang, Hua; Liu, Ying; Li, Guang-Xuan; Li, Qingwang; Hu, Jianhong

doi:10.1007/s10616-013-9584-0

Cited by 15 publications

(11 citation statements)

References 35 publications

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“…It has been shown that mouse SYCP1 functions in synaptonemal complex assembly, meiotic recombination, and XY body formation . The expression of SYCP3 is related to the differentiation of the spermatogonial stem cells into haploid male germ cells . XRN2 is a 5′‐3′ exoribonuclease that is essential for embryogenesis, larval development, and fertility of C. elegance .…”

Section: Discussionmentioning

confidence: 99%

“…37 The expression of SYCP3 is related to the differentiation of the spermatogonial stem cells into haploid male germ cells. 38 XRN2 is a 5′-3′ exoribonuclease that is essential for embryogenesis, larval development, and fertility of C. elegance. 39 Spermatid Perinuclear RNA Binding Protein (STRBP) is involved in spermatogenesis process and thus male fertility.…”

Section: Discussionmentioning

confidence: 99%

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In silico analysis of single‐cell RNA sequencing data from 3 and 7 days old mouse spermatogonial stem cells to identify their differentially expressed genes and transcriptional regulators

Sisakhtnezhad

2018

J of Cellular Biochemistry

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Spermatogonial stem cells (SSCs), which are at the basis of spermatogenesis process, are valuable cells with different applications in biotechnology and regenerative medicine. Understanding the molecular basis of SSC self-renewal and differentiation at various developmental stages of the male organism is crucial to find key factors in the SSCs fate and function. Therefore, this study was aimed to use single-cell RNA-sequencing dataset analysis for identification of differentially expressed genes (DEGs) and their regulators in 3 and 7 days old mouse-derived single SSCs (mSSCs). Results showed 68 upregulated and 203 downregulated genes in 7 days old mouse-derived SSCs compared to 3 days old mSSCs, which were associated with 1493 and 3077 biological processes, respectively. It also found that DAZL, FKBP6, PAIP2, DDX4, H3F3B, TEX15, XRN2, MAEL, and SOD1 are important factors with the higher gene expression pattern, which may be pivotal for mSSCs fate and function during development of germ cells. Moreover, NR3C1, RXRA, NCOA, ESR1, PML, ATF2, BMI1, POU5F1, and CHD1 were the main central regulators for the upregulated DEGs, while HNF1A, C/EBPα, and NFATC1 were the master regulators for the downregulated DEGs. In this regard, two significant protein complexes were found in the protein-protein interactions network for the upregulated DEGs regulators. Furthermore, 24 protein kinases detected upstream of the main central regulators of DEGs. In conclusion, this study presents DEGs and their transcriptional regulators that are crucial for inducing and regulating SSCs commitment during development, and for developing efficient protocols to identify and isolate SSCs for different applications.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

In silico analysis of single‐cell RNA sequencing data from 3 and 7 days old mouse spermatogonial stem cells to identify their differentially expressed genes and transcriptional regulators

Sisakhtnezhad

2018

J of Cellular Biochemistry

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“…Cells were cultured at 37.1 °C under an atmosphere of 95% humidified air with 5% CO 2 . To induce differentiation, the cells were cultured in medium containing retinoic acid (Sigma Chemical Company, St. Louis, MO, USA) and other growth factors described in literature14. To examine the effect of IL-6, the cells were cultured in medium containing 0, 10, 50 and 100 ng/ml IL-6.…”

Section: Methodsmentioning

confidence: 99%

IL-6 mediates differentiation disorder during spermatogenesis in obesity-associated inflammation by affecting the expression of Zfp637 through the SOCS3/STAT3 pathway

Huang

Miao

Zhang

et al. 2016

Sci Rep

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Zfp637 is a recently identified zinc finger protein, and its functions remain largely unknown. Here, we innovatively demonstrate the effects of Zfp637 on the differentiation of mouse spermatogonia and on its downstream target gene SOX2 in vitro. Obesity has been recognized as a chronic inflammatory disease that leads to decreased sexual function and sexual development disorders. We observed higher levels of IL-6 in serum and testis homogenates from obese mice compared with control mice. We also demonstrated that high levels of IL-6 inhibited Zfp637 expression, and we elucidated the underlying mechanisms. SOCS3 overexpression and STAT3 phosphorylation inhibitor (AG490) were used to investigate the function of the SOCS3/STAT3 pathway during this process. Our results showed that exposure of mouse spermatogonial cells to high levels of IL-6 inhibited Zfp637 expression by increasing SOCS3 expression and inhibiting the phosphorylation of STAT3, further reducing cellular differentiation. Consistent with the in vitro results, we observed increasing expression levels of SOCS3 and SOX2, but a reduction of Zfp637 expression, in obese mouse testes. In conclusion, Zfp637 plays a crucial role in spermatogenesis by downregulating SOX2 expression, and IL-6 can decrease the expression of Zfp637 through the SOCS3/STAT3 signaling pathway.

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“…More recently, different approaches have been proposed that could direct the differentiation of SSCs all the way to haploid male germ cells. Using isolated 6-day-old mouse SSCs, production of haploid germ cells was achieved by culturing testicular cells in 10% FBS-supplemented media for 3 days, changing the media into a retinoic acid (RA)-enriched medium to induce differentiation for 2 days, and followed by changing the media again to the first medium for 6-8 days [107].…”

Section: Two-dimensional Culture Of Testis Cell Suspensionsmentioning

confidence: 99%

Spermatogonial Stem Cells for In Vitro Spermatogenesis and In Vivo Restoration of Fertility

Ibtisham

Honaramooz

2020

Cells

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Spermatogonial stem cells (SSCs) are the only adult stem cells capable of passing genes onto the next generation. SSCs also have the potential to provide important knowledge about stem cells in general and to offer critical in vitro and in vivo applications in assisted reproductive technologies. After century-long research, proof-of-principle culture systems have been introduced to support the in vitro differentiation of SSCs from rodent models into haploid male germ cells. Despite recent progress in organotypic testicular tissue culture and two-dimensional or three-dimensional cell culture systems, to achieve complete in vitro spermatogenesis (IVS) using non-rodent species remains challenging. Successful in vitro production of human haploid male germ cells will foster hopes of preserving the fertility potential of prepubertal cancer patients who frequently face infertility due to the gonadotoxic side-effects of cancer treatment. Moreover, the development of optimal systems for IVS would allow designing experiments that are otherwise difficult or impossible to be performed directly in vivo, such as genetic manipulation of germ cells or correction of genetic disorders. This review outlines the recent progress in the use of SSCs for IVS and potential in vivo applications for the restoration of fertility.Cells 2020, 9, 745 2 of 31 manipulation. Moreover, optimal conditions for the in vitro culture and propagation of SSCs are also needed to help boost the potential therapeutic application of SSCs in fertility preservation or restoration. Long-term culture of rodent SSCs is well demonstrated; however, relatively few in vitro culture conditions have been examined for primate SSCs [2]. Any potential in vivo application of cultured human SSCs, such as in restoration of fertility, requires extensive studies to ensure their safety and efficacy. The establishment of efficient culture conditions for human SSCs also necessitates the availability of proper animal models, for instance, xenotransplantation techniques to assess the quality and quantity of such cultured SSCs; these assays have so far been used sporadically for testing primate SSCs [4].Spermatogenesis is a complex process of germ cell proliferation and differentiation that requires extensive interactions among different cell types, hormones, growth factors, and various other signals, making it difficult to be replicated in vitro. There are several objectives for establishing an optimal and efficient culture system to recapitulate the process of germ cell development in vitro. This includes the study of basic requirements of male germ cell development, proliferation, differentiation, and production of haploid germ cells in a controlled in vitro environment. Additionally, such culture systems could be potentially used to produce haploid male germ cells from undifferentiated germ cells isolated from the testis of infertile adult patients and/or testicular biopsies collected from prepubertal cancer patients before undergoing gonadotoxic treatment. The establishment of ...

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Differentiation of spermatogonial stem cell-like cells from murine testicular tissue into haploid male germ cells in vitro

Cited by 15 publications

References 35 publications

In silico analysis of single‐cell RNA sequencing data from 3 and 7 days old mouse spermatogonial stem cells to identify their differentially expressed genes and transcriptional regulators

In silico analysis of single‐cell RNA sequencing data from 3 and 7 days old mouse spermatogonial stem cells to identify their differentially expressed genes and transcriptional regulators

IL-6 mediates differentiation disorder during spermatogenesis in obesity-associated inflammation by affecting the expression of Zfp637 through the SOCS3/STAT3 pathway

Spermatogonial Stem Cells for In Vitro Spermatogenesis and In Vivo Restoration of Fertility

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