Effective isolation of Sertoli cells from New Zealand rabbit testis

Zhu, Wenyao; Yang, De-Cai; Jiang, Yu; Cai, Ningning; Yang, Rui; Zhang, Xueming

doi:10.5455/javar.2021.h505

Cited by 3 publications

(2 citation statements)

References 19 publications

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“…It will contribute to the elimination of the lack of knowledge in this field by considering the current and deficiencies. The findings of this study were carried out in scientific studies on the male genital system in laboratory animals and other animal species (Castellini et al, 2022; Emmanuel et al, 2019), artificial insemination studies (Ata et al, 2018; Dimitrova et al, 2009; Lukac et al, 2009) is thought to contribute to reproductive system diseases and operations in rabbits (Harcourt‐Brown, 2017;Zhu et al, 2018; Reineking et al, 2019; Zhu et al, 2021).…”

Section: Discussionmentioning

confidence: 99%

Investigation of male genital system anatomy in the New Zealand rabbit (Oryctolagus cuniculus L.)

Bozbiyik

Doğan

2022

Anat Histol Embryol

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Rabbits are frequently used in research because of their human-like biological structure. In the present study, 10 male rabbits were preferred. Arterial vascularization of the genital system organs were dissected. Morphometric and macroanatomical results of these organs were taken, and some organs were examined by magnetic resonance imaging (MRI). In this study, right testis length was measured at 67.24 ± 5.36 mm and left testis length was 62.25 ± 6.55 mm in New Zealand rabbits.Testicular weight was 10.21 ± 1.17 g on the right and 9.90 ± 1.15 g on the left. Right caput epididymidis 0.68 ± 0.08 g, left caput epididymidis 0.80 ± 0.13 g; right corpus epididymidis 0.14 ± 0.03 left corpus epididymidis 0.15 ± 0.03 g; right cauda epididymidis 1.87 ± 0.26 g left cauda epididymidis 1.90 ± 0.31 g. In the present study, right ductus deferens weight was 0.30 ± 0.29 g, and left ductus deferens weight was 0.25 ± 0.03 g; the ampulla ductus deferentis weight was 0.35 ± 0.16 g on the right side and 0.37 ± 0.16 g on the left side. The length of the vesicular gland glandula vesicularis was calculated as 18.54 ± 0.35 mm on the right and 17.55 ± 0.59 mm on the left. The width of the vesicular gland was measured as 16.54 ± 0.28 mm on the right and 16.70 ± 0.45 mm on the left. Vesicular gland weight was 0.91 ± 0.07 g on the right side and 0.96 ± 0.09 g on the left side. The prostate length was recorded as 11.68 ± 2.01 mm and width as 13.02 ± 1.38 mm. The length of the bulbourethral gland was 13.36 ± 2.00 mm on the right side and 12.39 ± 1.21 mm on the left. Its width was recorded as 6.73 ± 0.98 mm on the right and 5.80 ± 0.90 mm on the left. Respectively, it weighed 0.40 ± 0.20 g on the right and 0.44 ± 0.24 g on the left. The prostate consisted of three parts in rabbits: the proprostate, the corpus prostate and the paraprostate. The bulbourethral gland was a green lentil-sized gland found in pairs. It is thought that the obtained results will contribute to the scientific researches on male genital system in laboratory animals and other animal species, artificial insemination studies, reproductive system diseases and operations in rabbits.

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Section: Discussionmentioning

confidence: 99%

Investigation of male genital system anatomy in the New Zealand rabbit (Oryctolagus cuniculus L.)

Bozbiyik

Doğan

2022

Anat Histol Embryol

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“…The enriched cells were all positive for the markers using antibodies against GATA4 (ab84593, Abcam), SOX9 (ab185966, Abcam), and ABP (bs‐2410R, Bioss Inc.) as previously described (Figure S1A–C). 26 …”

Section: Methodsmentioning

confidence: 99%

The role of hepatitis B virus surface protein in inducing Sertoli cell ferroptosis

Pan,

Kong,

et al. 2023

Andrology

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BackgroundsHepatitis B virus infection could result in male infertility with sperm defects and dysfunction. Sertoli cells are essential for testis function and play a crucial role in spermatogenesis. Sertoli cell death contributes to spermatogenesis impairment, leading to poor sperm quality. Ferroptosis has been implicated as a mechanism of Sertoli cell death. The issue in studying the relationship between hepatitis B virus and Sertoli cell ferroptosis has not yet been addressed.ObjectivesTo explore the mechanisms underlying ferroptosis in hepatitis B virus‐exposed Sertoli cells.Materials and methodsHuman Sertoli cells were treated in vitro with levels of 25, 50, and 100 μg/mL of hepatitis B virus surface protein (HBs). Cell viability and levels of glutathione, malondialdehyde, cellular ferrous ion (Fe2+), lipid peroxidation, and N6‐methyladenosine in Sertoli cells were detected. The level of glutathione peroxidase 4, transferrin receptor 1, ferritin heavy chain, tripartite motif (TRIM) 37, methyltransferase like 3, and insulin‐like growth factor 2 mRNA binding protein 2 was examined. Cell transfection was carried out to alter expression of ferroptosis‐related proteins. qPCR and immunoblotting were performed to measure protein expression level. Immunoprecipitation was applied to determine the protein and protein‐RNA interaction. Luminescence analysis was performed to identify the target of methyltransferase like 3.ResultsHBs exposure triggered ferroptosis featured with increased intracellular Fe2+ ion, reduced cell viability and expression of glutathione peroxidase 4 in Sertoli cells. HBs treatment significantly increased TRIM37 expression, which suppressed glutathione peroxidase 4 expression through ubiquitination. TRIM37 silencing attenuated the effect of HBs exposure‐regulated cell viability and ferroptosis. HBs upregulated N6‐methyladenosine modification in TRIM37 3′‐UTR by increasing methyltransferase like 3 expression. The binding of N6‐methyladenosine reader insulin‐like growth factor 2 mRNA binding protein 2 and TRIM37 3′‐UTR enhanced the stability of TRIM37 mRNA.ConclusionHBs can decrease human Sertoli cell viability by promoting ferroptosis induced by the loss of glutathione peroxidase 4 activity through TRIM37‐mediated ubiquitination of glutathione peroxidase 4. The findings highlight the role of TRIM37/glutathione peroxidase 4 signaling responsible for ferroptosis regulation in hepatitis B virus‐infected Sertoli cells.

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Co-Culture of Cryopreserved Healthy Sertoli Cells with Testicular Tissue of Non-Obstructive Azoospermia (NOA) Patients in Culture Media Containing Follicle-Stimulating Hormone (FSH)/Testosterone Has No Advantage in Germ Cell Maturation

Aydos

Yükselten

Özkan

et al. 2023

JCM

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Different cell culture conditions and techniques have been used to mature spermatogenic cells to increase the success of in vitro fertilization. Sertoli cells (SCs) are essential in maintaining spermatogenesis and FSH stimulation exerts its effect through direct or indirect actions on SCs. The effectiveness of FSH and testosterone added to the co-culture has been demonstrated in other studies to provide microenvironment conditions of the testicular niche and to contribute to the maturation and meiotic progression of spermatogonial stem cells (SSCs). In the present study, we investigated whether co-culture of healthy SCs with the patient’s testicular tissue in the medium supplemented with FSH/testosterone provides an advantage in the differentiation and maturation of germ cells in NOA cases (N = 34). In men with obstructive azoospermia (N = 12), healthy SCs from testicular biopsies were identified and purified, then cryopreserved. The characterization of healthy SCs was done by flow cytometry (FC) and immunohistochemistry using antibodies specific for GATA4 and vimentin. FITC-conjugated annexin V/PI staining and the MTT assay were performed to compare the viability and proliferation of SCs before and after freezing. In annexin V staining, no difference was found in percentages of live and apoptotic SCs, and MTT showed that cryopreservation did not inhibit SC proliferation compared to the pre-freezing state. Then, tissue samples from NOA patients were processed in two separate environments containing FSH/testosterone and FSH/testosterone plus co-culture with thawed healthy SCs for 7 days. FC was used to measure 7th-day levels of specific markers expressed in spermatogonia (VASA), meiotic cells (CREM), and post-meiotic cells (protamine-2 and acrosin). VASA and acrosin basal levels were found to be lower in infertile patients compared to the OA group (8.2% vs. 30.6% and 12.8% vs. 30.5%, respectively; p < 0.05). Compared to pre-treatment measurements, on the 7th day in the FSH/testosterone environment, CREM levels increased by 58.8% and acrosin levels increased by 195.5% (p < 0.05). Similarly, in medium co-culture with healthy SCs, by day 7, CREM and acrosin levels increased to 92.2% and 204.8%, respectively (p < 0.05). Although VASA and protamine levels increased in both groups, they did not reach a significant level. No significant difference was found between the day 7 increase rates of CREM, VASA, acrosin and protamine-2 in either FSH/testosterone-containing medium or in medium additionally co-cultured with healthy SCs (58.8% vs. 92.2%, 120.6% vs. 79.4%, 195.5% vs. 204.8%, and 232.3% vs. 198.4%, respectively; p > 0.05). Our results suggest that the presence of the patient’s own SCs for maturation of germ cells in the culture medium supplemented with FSH and testosterone is sufficient, and co-culture with healthy SCs does not have an additional advantage. In addition, the freezing–thawing process would not impair the viability and proliferation of SCs.

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Effective isolation of Sertoli cells from New Zealand rabbit testis

Cited by 3 publications

References 19 publications

Investigation of male genital system anatomy in the New Zealand rabbit (Oryctolagus cuniculus L.)

Investigation of male genital system anatomy in the New Zealand rabbit (Oryctolagus cuniculus L.)

The role of hepatitis B virus surface protein in inducing Sertoli cell ferroptosis

Co-Culture of Cryopreserved Healthy Sertoli Cells with Testicular Tissue of Non-Obstructive Azoospermia (NOA) Patients in Culture Media Containing Follicle-Stimulating Hormone (FSH)/Testosterone Has No Advantage in Germ Cell Maturation

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