Human Sperm Bioassay for Reprotoxicity Testing in Embryo Culture Media: Some Practical Considerations in Reducing the Assay Time

Hossain, Amjad; Aryal, Subhash; Osuampke, Collin; Phelps, John Y.

doi:10.1155/2010/136898

Cited by 9 publications

(4 citation statements)

References 23 publications

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“…The assessment of sperm motility was carried out according to the WHO protocol. 24,25 Epididymal Sperm Morphology. Following previous observations, a small amount of sperm suspension was smeared onto a slide by using a pipet and then fixed with methanol.…”

Section: ■ Materials and Methodsmentioning

confidence: 99%

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Toxic Effects and Possible Mechanisms of Deoxynivalenol Exposure on Sperm and Testicular Damage in BALB/c Mice

Yang

Wang

Guo

et al. 2019

J. Agric. Food Chem.

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Deoxynivalenol (DON, vomitoxin) is the most common mycotoxin in cereals and grains. DON contamination can cause a serious health threat to humans and farm animals. DON has been reported to exert significant toxicity effects on the male reproductive system. However, the causes and mechanisms underlying efforts of DON on sperm and testicular damage remain largely unclear. In the present study, we thoroughly investigated this issue. Eighty male BALB/c mice were randomly divided into a control group (n = 40) and DON treatment group (2.4 mg/kg of body weight, n = 40). The ratio of testes and seminal vesicle to body, sperm survival and motility, and morphology of sperm and testis were observed in DON-treated and control mice. In addition, the concentrations of reactive oxygen species (ROS) and malondialdehyde (MDA), the activities of superoxide dismutase (SOD) and glutathione (GSH), and also the expression levels of JNK/c-Jun signaling and apoptotic factors such as caspase-3, caspase-8, caspase-9, Bim, and Bid were analyzed and compared between the two groups. The results demonstrated that a single topical application of DON significantly increased the percentage of abnormal sperm and decreased the motility of sperm, indicating the sperms are damaged by DON. Additionally, the reduced relative body weight of testis and severe destruction of testicular morphology were observed. Moreover, the increased levels of ROS and MDA levels and decreased activities of SOD and GSH were found in testicular tissues, suggesting that oxidative stress is induced by DON treatment. Furthermore, DON upregulated the expression of stress-induced JNK/c-Jun signaling pathway proteins as well as JNK/c-Jun phosphorylation proteins. In addition, DON could enhance testicular apoptosis by increasing expression levels of apoptotic genes including Bim, cytochrome c, caspase 3, caspase 8, and caspase 9. These results suggest that DON exposure can cause sperm damage, oxidative stress, testicular apoptosis, and phosphorylation of JNK/c-Jun signaling pathway. The underlying mechanisms may be that DON induces sperm damage by exacerbating oxidative stress-mediated testicular apoptosis via JNK/c-Jun signaling pathway.

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Section: ■ Materials and Methodsmentioning

confidence: 99%

“…The parameters for sperm motility were the percentage of motile sperm and the percentage of progressive motile sperm. The assessment of sperm motility was carried out according to the WHO protocol. , …”

Section: Methodsmentioning

confidence: 99%

Toxic Effects and Possible Mechanisms of Deoxynivalenol Exposure on Sperm and Testicular Damage in BALB/c Mice

Yang

Wang

Guo

et al. 2019

J. Agric. Food Chem.

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“…Human sperm bioassays are highly sensitive when both motility and the quality of sperm motility (motility grade) are taken into consideration [7]. The American Association of Bioanalysts (AAB) chose an assay time of 48 hours for identifying the quality of ART culture media, we used 48 hours as an endpoint to be consistent with this guidance, and assayed motility percent and rapid progression percent, to sensitively quantify causal association of sperm performance to handling medium.…”

Section: Global Journal Of Reproductive Medicinementioning

confidence: 99%

Optimization of Sperm Culture Conditions for Human Assisted Reproductive Technologies

Bormann¹

2018

GJORM

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Optimizing sperm performance can be a differentiating factor in clinical pregnancy rates for assisted reproductive technology (ART) laboratories. Minimizing cellular stress during routine sperm processing by maintaining a precise and stable environment is of great importance in the field. In commercial media, pH of the external cellular culture environment (pHe) is typically maintained at 7.2-7.4 by various buffering reagents, while the internal pH of the cell (pHi) depends on available lactate and amino acids. Combining buffers in solution has been previously shown to be valuable for stabilizing internal and external pH for various biological systems. We hypothesized that a dual-buffer culture medium might improve sperm performance for ART. Here we demonstrate superior performance of a commercially available dual-buffer solution of HEPES and MOPS: Multipurpose Handling Medium (MHM, Irvine Scientific). Significantly better performance, assessed at 8 and 48 hours, and measured as: sperm viability, total motility, and rapid forward progression, was observed for MHM over single buffer controls.

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“…Prostaglandin E synthase (PGES) synthesizes PGE 2 from PGH 2 . The COX-2/PGE 2 pathway has been well studied in regards to chronic inflammation and cancer development [ 12 , 13 ]. Bioinformatics analysis of urothelial cells treated with antiproliferative factor (APF), has implicated COX-2 as being involved in IC/BPS however, the extent is unknown [ 14 ].…”

Section: Introductionmentioning

confidence: 99%

Impaired Expression of Prostaglandin E2 (PGE2) Synthesis and Degradation Enzymes during Differentiation of Immortalized Urothelial Cells from Patients with Interstitial Cystitis/Painful Bladder Syndrome

2015

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PurposeThe differentiated superficial cells of the urothelium restrict urine flow into the bladder wall. We have demonstrated that urothelial cells isolated from bladders of patients with interstitial cystitis/painful bladder syndrome (IC/PBS) fail to release PGE2 in response to tryptase. This study examines the expression of PGE2 synthesis and degradation enzymes in urothelial cells during differentiation.Materials and MethodsWe measured immunoprotein expression of cyclooxygenase-2 (COX-2), prostaglandin E2 synthase (PGES) and 15-hydroxyprostaglandin dehydrogenase (PGDH) in human urothelial cells and in immortalized urothelial cells isolated from the bladders of IC/PBS patients or normal subjects during stratification and differentiation produced by increased calcium and fetal bovine serum (Ca/FBS) in the culture medium for 1, 3 and 7 days.ResultsPGES immunoprotein expression increased during differentiation in normal and IC/PBS urothelial cells. COX-2 expression also increased in cells from normal patients following differentiation. Remarkably, no COX-2 expression was detectable in urothelial cells isolated from 3 out of 4 IC/PBS patients. PGDH immunoprotein expression decreased in normal cells after 1 and 3 days of Ca/FBS addition, but returned to normal after 7 days. PGDH expression was unchanged during differentiation at 1 and 3 days, but was more than 2-fold higher at 7 days compared to day 0 in the IC/PBS cells. Urothelial cells isolated from IC/PBS patients demonstrated no PGE2 release in response to tryptase under any of the experimental conditions studied.ConclusionsTaken together, our results indicate that PGE2 release is compromised during stratification and differentiation in IC/PBS urothelium and may contribute to impaired barrier function.

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Human Sperm Bioassay for Reprotoxicity Testing in Embryo Culture Media: Some Practical Considerations in Reducing the Assay Time

Cited by 9 publications

References 23 publications

Toxic Effects and Possible Mechanisms of Deoxynivalenol Exposure on Sperm and Testicular Damage in BALB/c Mice

Toxic Effects and Possible Mechanisms of Deoxynivalenol Exposure on Sperm and Testicular Damage in BALB/c Mice

Optimization of Sperm Culture Conditions for Human Assisted Reproductive Technologies

Impaired Expression of Prostaglandin E2 (PGE2) Synthesis and Degradation Enzymes during Differentiation of Immortalized Urothelial Cells from Patients with Interstitial Cystitis/Painful Bladder Syndrome

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