陳煜、*鄭兆珉、*黄翰威、†Koji Matsuura
林口長庚紀念醫院 外科部泌尿科 長庚大學；*清華大學生物醫學工程研究所；
†岡山大學工學部生物醫學工程系
Relationship between porcine sperm motility and
Sperm enzymatic activity using paper-based devices-0
Yu Chen, Chao-Min Cheng*, Han-Wei Huang*, Koji Matsuura†
Divisions of Urology, Department of Surgery, Chang Gung Memorial Hospital and Chang Gung University
*Institute of Biomedical Engineering, National Tsing Hua University
†Department of Biomedical Engineering, Faculty of Engineering, Okayama University
Purpose: 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.
Materials and Methods: 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 value of area mean 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).
Results: 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.
Conclusion: Based on this inhibitor study, sperm motility can be estimated using our paper-based MTT-assay.