Evaluation of cockerel spermatozoa viability and motility by a novel enzyme based cell viability assay

Abstract

1. The results of spermatozoa assessment by the WST-8 (2-[2-methoxy-4-nitrophenyl]-3-[4-nitrophenyl]-5-[2,4-disulfophenyl]-2H-tetrazolium, monosodium salt) assay, flow cytometry (FC) or computer-assisted sperm analysis (CASA) were compared. 2. Different live/killed ratios of cockerel semen were serially diluted to 120, 60, and 30 × 10 cells/ml, and each sample was analysed by (1) WST-8 assay at 0, 10, 20, 30, 40, 50, 60 min, (2) viability with FC, and (3) motility with CASA. 3. The WST-8 reduction rate was closely correlated with spermatozoa viability and motility. The optimal semen concentration for the WST-8 assay was 120 × 10 cells/ml, and the standard curves for spermatozoa viability and motility predictions, respectively, were y = 162.8x + 104.96 (R = 0.9594) after 60 min of incubation and y = 225.09x + 96.299 (R = 0.8475) after 40 min of incubation. 4. It was concluded that the WST-8 assay is useful for the practical evaluation of cockerel spermatozoa viability and motility. Compared to FC and CASA, the WST-8 assay does not require expensive and complex instrumentation in the lab. Furthermore, one well of the WST-8 reaction can be used to predict spermatozoa viability and motility at the same time, which all lead it to be efficient and economical for semen quality assessment.

Molecular Mechanism of Apoptotic Cell Death in Cyanidin-3-glucoside-induced Cytotoxic Potential on Human Liver Carcinoma (HepG2) Cell Line

“…A similar dose-based growth reduction has been reported when C3gc was tested against H661 lung cancer cells and MDA-MB-453 breast cancer cells. 27 Following the disruption of the cytoplasmic membrane and cell necrosis, damaged or stressed cells might leak cytoplasmic LDH and other molecules into the medium. 28 Consequently, it's probable that the C3gc cytotoxic action wasn't dependent on the process of necrosis-induced cell death.…”

In vitro Effects of the Methanolic Leaf Extract of Otholobium fruticans in Murine B16 Melanoma Cells: Implications for the Treatment of Skin Hyperpigmentation

“…The 3-(4, 5-dimethyl thiazol-2-yl)-2, 5-diphenyl tetrazolium bromide (MTT) is a colorimetric assay used to determine cell viability based on the ability of succinate dehydrogenase enzymes in the mitochondria of live cells to metabolize yellow MTT to an insoluble purple colour formazan product which can be measured spectrophotometrically between 500 and 600 nm wavelengths [7,8,9]. Briefly, cells were grown at 6.0 x 10 4 cells/mL in 12-well culture plates and treated with plant extracts at various concentrations for 72 hrs.…”