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Nasr-Esfahani, Mohammad Hossein; Aboutorabi, Roshanak; Esfandiari, Ebrahim; Mardani, Mohammad

doi:10.1023/a:1020310503143

Cited by 41 publications

(8 citation statements)

References 14 publications

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“…Berg and Nasr-Esfahani et al . reported that MTT formazan is accumulated at the midpiece, and recognized as granules under optical microscope5051. Redox reactions to reduce MTT catalyzed by redox enzymes occur at midpiece mitochondria and some of these redox reactions would not couple with sperm motility.…”

Section: Resultsmentioning

confidence: 99%

Relationship between Porcine Sperm Motility and Sperm Enzymatic Activity using Paper-based Devices

Matsuura

Huang

Chen

et al. 2017

Sci Rep

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Mammalian sperm motility has traditionally been analyzed to determine fertility using computer-assisted semen analysis (CASA) systems. To develop low-cost and robust male fertility diagnostics, we created a paper-based MTT assay and used it to estimate motile sperm concentration. When porcine sperm motility was inhibited using sperm enzyme inhibitors for sperm enzymes related to mitochondrial activity and glycolysis, we simultaneously recorded sperm motility and enzymatic reactivity using a portable motility analysis system (iSperm) and a paper-based MTT assay, respectively. When using our paper-based MTT-assay, we calculated the area mean value signal intensity (AMV) to evaluate enzymatic reactivity. Both sperm motility and AMV decreased following treatment with iodoacetamide (IODO) and 3-bromopyruvic acid (3BP), both of which are inhibitors of glycolytic enzymes including glyceraldehyde-3-phosphate dehydrogenase (GAPDH). We found a correlation between recorded motility using iSperm and AMV from our paper-based assay (P < 0.05), suggesting that a sperm-related enzymatic reaction is involved in sperm motility. Under this protocol, MTT reduction was coupled with catalysis of GAPDH and was promoted by electron transfer from NADH. Based on this inhibitor study, sperm motility can be estimated using our paper-based MTT-assay.

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Section: Resultsmentioning

confidence: 99%

Relationship between Porcine Sperm Motility and Sperm Enzymatic Activity using Paper-based Devices

Matsuura

Huang

Chen

et al. 2017

Sci Rep

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“…Previous studies using the MTT assay have calculated values of 14.8 and 14.6% for intra‐ and inter‐assay CVs respectively [12]. Nasr‐Esfahani et al [31] adapted the MTT assay for determining the viability of sperm and found 9 and 7% for intra‐ and inter‐ assay CVs respectively. Therefore the values in the present study of 7 and 6.6% for intra‐ and inter‐assay CVs were regarded as credible for determining viable cell number for the MLTC‐1 cell line.…”

Section: Resultsmentioning

confidence: 99%

Development of an in vitro reproductive screening assay for novel pharmaceutical compounds

Edwards¹,

Markovic²,

Matisons³

et al. 2008

Biotech and App Biochem

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An in vitro reproductive cell-based toxicity assay was developed using MLTC-1 (murine Leydig tumour cell line) in order to examine the reproductive toxicity of two novel nanopharmaceutical compounds, namely ethylene glycol mono allyl ether and poly(ethylene glycol) octa-functionalized polyhedral oligomeric silsesquioxane. Three commonly used cytotoxicity assays, namely the MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide], MTS [3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium] and Crystal Violet assays, were compared, and the MTT assay proved to be the most accurate and reproducible for the MLTC-1 cell line. The doubling rate of the MLTC-1 cells was 30+/-3.5 h and the optimal seeding density for the MTT assay was 20000 cells per well, and the optimized MTT assay utilized a 4 h cell adherence followed by incubation with 0.5 mg/ml MTT for 1 h. The intra- and inter-assay CV (coefficient of variation) values were 12.3 and 11% respectively. MLTC-1 cells only produce the reproductive hormone progesterone in response to hCG (human chorionic gonadotropin), which stimulated progesterone production dose-dependently from 0 to 100 m.i.u. (milliinternational units)/ml (2706+/-1118 ng/ml). H(2)O(2) as a negative control killed 100% of cells at 1000 microg/ml. The two nanopharmaceutical compounds were cytotoxic at concentrations > or =0.1 microg/ml, but hCG decreased cytotoxicity to > or =1000 microg/ml (P<0.001). hCG-stimulated progesterone synthesis afforded some protection against the cytotoxic effects of the two novel nanotechnology compounds; therefore doses < or =100 microg/ml and an exposure period of 1 h would be recommended for testing in in vivo animal reproductive assays.

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“…Subsequently, sperm evaluations were performed by applying semen onto the paper-based device treated with yellow MTT. Subsequently, catalytic sperm redox enzymes convert yellow MTT to the purple formazan metabolite [77][78][79]. Hence, the development of purple colour (8-bit grey scale converted) verifies enzyme activity and sperm motility.…”

Section: Paper-based Devices For Semen Diagnosismentioning

confidence: 99%

Recent advances in microfluidic diagnostic and treatment systems for assisted reproductive technologies in developing countries

Matsuura¹,

Nishina²,

Asano³

2017

Point-of-Care Diagnostics - New Progresses and Perspectives

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Abstract: Sperm MTT viability assay can be used as a diagnostic test for discrimination of viable sperms from sperm population.

Cited by 41 publications

References 14 publications

Relationship between Porcine Sperm Motility and Sperm Enzymatic Activity using Paper-based Devices

Relationship between Porcine Sperm Motility and Sperm Enzymatic Activity using Paper-based Devices

Development of an in vitro reproductive screening assay for novel pharmaceutical compounds

Recent advances in microfluidic diagnostic and treatment systems for assisted reproductive technologies in developing countries

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