Isolation of Sertoli Cells and Peritubular Cells from Rat Testes

Bhushan, Sudhanshu; Aslani, Ferial; Zhang, Zhengguo; Sebastian, Tim; Elsässer, Hans–Peter; Klug, Jörg

doi:10.3791/53389

Cited by 33 publications

(37 citation statements)

References 31 publications

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“…The FSH increases the proliferation of Sertoli cells, which ultimately increases the protection of the spermatogenic lineage and provides the nutrients they need. In addition, Sertoli cells release water into the seminiferous tubules and cause a collection of toxic materials from around the spermatogenic lineage, and they also interfere with the maintenance of the size and weight of the testicles (Bhushan et al, ). Sertoli cells with the help of the proliferation of the spermatogenic lineage, increasing in thickness of the tubes, and their diameters, which ultimately decreases the volume of the interstitial tissue by increasing the volume and thickness of the tubes, also an indicator of testicular activity (Rigi Manesh et al, ).…”

Section: Discussionmentioning

confidence: 99%

Nasturtium Officinale L. hydroalcoholic extract improved oxymetholone‐induced oxidative injury in mouse testis and sperm parameters

Bazm

Khazaei

et al. 2019

Andrologia

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Testicular tissue and sex hormones are sensitive to the anabolic steroids (oxymetholone/OM) due to increased free radical damage and hormonal changes. The Nasturtium officinale L. have various antioxidant compounds. The aim of the present study was to investigate N. officinale effect on OM-induced oxidative injury in mouse testis and sperm parameters. Thirty BALB/c mice were divided into five groups, including control, OM (5 ml/kg) and three N. officinale doses (25, 50 and 100 mg/ kg) + OM. At the end of the study (40 days), serum luteinising hormone (LH), folliclestimulating hormone (FSH), testosterone, nitric oxide (NO) levels, ferric reducing ability of power (FRAP) and testis stereological factors were measured. The sperm parameters were evaluated. Liquid chromatography-electrospray ionisation-tandem mass spectrometry (LC-ESI/MS) analysis was yielded a fingerprint of N. officinale phenolic constituents. 100 mg/kg of N. officinale extract significantly reduced the serum level of testosterone and a significant increase in LH and FSH in comparison with the control group. This dose also significantly improved the stereological factors and sperm parameters. 50 and 100 mg/kg of N. officinale extract significantly increased the testis tissue FRAP levels, and 100 doses reduced the serum levels of NO.Fourteen compounds and 34 peaks were identified in the extract with LC-ESI/MS. Nasturtium officinale extract has protective effects against testicular toxicity caused

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Section: Discussionmentioning

confidence: 99%

Nasturtium Officinale L. hydroalcoholic extract improved oxymetholone‐induced oxidative injury in mouse testis and sperm parameters

Bazm

Khazaei

et al. 2019

Andrologia

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“…Primary Sertoli cells (SC), peritubular cells (PTC), and Leydig cells (LC) were isolated according to a reported procedure (23,24).…”

Section: Isolation Of Testicular Cellsmentioning

confidence: 99%

Characterization of the Micro-Environment of the Testis that Shapes the Phenotype and Function of Testicular Macrophages

Wang

Fijak

Hossain

et al. 2017

The Journal of Immunology

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105

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Macrophages are important in the activation of innate immune responses and in a tissue-specific manner in the maintenance of organ homeostasis. Testicular macrophages (TM), which reside in the testicular interstitial space, comprise the largest leukocyte population in the testes and are assumed to play a relevant function in maintaining testicular immune privilege. Numerous studies have indicated that the interstitial fluid (IF) surrounding the TM has immunosuppressive properties, which may influence the phenotype of TM. However, the identity of the immunosuppressive molecules present in the IF is poorly characterized. We show that the rat testicular IF shifted GM-CSF–induced M1 toward the M2 macrophage phenotype. IF-polarized M2 macrophages mimic the properties of TM, such as increased expression of CD163, high secretion of IL-10, and low secretion of TNF-α. In addition, IF-polarized macrophages display immunoregulatory functions by inducing expansion of immunosuppressive regulatory T cells. We further found that corticosterone was the principal immunosuppressive molecule present in the IF and that the glucocorticoid receptor is needed for induction of the testis-specific phenotype of TM. In addition, TM locally produce small amounts of corticosterone, which suppresses the basal expression of inflammatory genes as a means to render TM refractory to inflammatory stimuli. Taken together, these results suggest that the corticosterone present in the testicular environment shapes the immunosuppressive function and phenotype of TM and that this steroid may play an important role in the establishment and sustenance of the immune privilege of the testis.

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“…35,36 Briefly, the testes from the supernatants were carefully removed, and the tubules were washed for 9 times with PBS to remove contaminating interstitial cells. 35,36 Briefly, the testes from the supernatants were carefully removed, and the tubules were washed for 9 times with PBS to remove contaminating interstitial cells.…”

Section: Cell Isolation and Culture Of Primary Scs And Tm4 Cell Linesmentioning

confidence: 99%

“…SCs were isolated as described previously. 35,36 Briefly, the testes from 2-week-old ICR male mice were collected and washed with PBS. The tunica-free testes were then incubated with 2.5 mg/mL trypsin (Gibco, Grand Island, NY, USA) and 10 μg/mL DNase I (Sigma, St. Louis, MO, USA) at 32°C for 5 minutes with gentle oscillation.…”

Section: Cell Isolation and Culture Of Primary Scs And Tm4 Cell Linesmentioning

confidence: 99%

Identification of programmed cell death 1 and its ligand in the testicular tissue of mice

Wang

Duan

et al. 2019

American J Rep Immunol

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Problem This study aims to determine the expression and localization of programmed cell death 1 (PD‐1) and programmed cell death 1 ligand 1 (PD‐L1) in the testes of mice at different developmental stages. Method of study By means of RT‐qPCR, Western blot and immunofluorescence, the expression and localization of PD‐1 and PD‐L1 were detected in the testicular tissues of mice at different postnatal times: P7, P14, P21, P28, P35, and adulthood. Meanwhile, the level of soluble PD‐L1 (sPD‐L1) was evaluated by ELISA in the testicular interstitial fluid (IF) of the adult mice, culture supernatants of TM4 cell lines (Sertoli cells lines), and primary Sertoli cells at P14. Results Pd‐1 mRNA levels were unexpectedly low. From P7 to P21, there was limited PD‐1 protein detected while PD‐1 was evident at P28 and afterward at significantly higher levels than at P14 and P21 (P < 0.05). Despite being found in the interstitial area at P7, P14, and P21, PD‐1 was also detected in the germ cells of the seminiferous tubules after P28. Pd‐l1 mRNA exhibited age‐related changes, peaking at P21, while PD‐L1 protein was constitutively expressed at any stage, specifically localized in the nucleus of Sertoli cells. Moreover, the level of sPD‐L1 in IF was significantly higher than that in the culture supernatants of both TM4 and primary Sertoli cells at P14. Conclusions PD‐1 and PD‐L1 were present in the testicular tissue of adult mice. The expression and localization of PD‐1 fluctuated with age, and PD‐1 was mainly localized to advanced germ cells, suggesting that it may play a role in spermiogenesis. PD‐L1 was constitutively expressed in the nucleus of Sertoli cells, which could secrete sPD‐L1 into the testicular interstitial space and thus may be involved in testicular immune privilege.

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Isolation of Sertoli Cells and Peritubular Cells from Rat Testes

Cited by 33 publications

References 31 publications

Nasturtium Officinale L. hydroalcoholic extract improved oxymetholone‐induced oxidative injury in mouse testis and sperm parameters

Nasturtium Officinale L. hydroalcoholic extract improved oxymetholone‐induced oxidative injury in mouse testis and sperm parameters

Characterization of the Micro-Environment of the Testis that Shapes the Phenotype and Function of Testicular Macrophages

Identification of programmed cell death 1 and its ligand in the testicular tissue of mice

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