Human Sperm Cells After Purification Using SCLB Can Be Stored at 4o, -20o, or -80oC Before Small RNA Isolation

Abstract

There have been many studies about pre-analysis for sperm RNA examination which compared sperm purificationmethods, RNA isolation methods, sequencing methods, and semen storage before analysis. However, there is a lack ofstudies that determine the ideal storage temperature after sperm cell purification before RNA analysis, especially small RNAanalysis. The aim of this study was to determine the preferred storage temperature for human sperm cells after spermpurification using Somatic Cell Lysis Buffer (SCLB) before sperm small ribonucleic acid (RNA) isolation and analysis. Thisstudy was a true laboratory experiment using the post-test only control group design. The samples were 13 fresh humansemen that has been purified using SCLB. The sperm cells were then diluted and divided into four aliquots with differenttreatments. The first aliquot that served as a control group was immediately purified while the other three aliquots were0 0 0 stored for seven days at different temperatures as follows: 4 C, -20 , and -80 C. After the small RNA isolation, RNA levelbetween each group was compared. Micro volume spectrophotometer measured RNA level. The median of small RNA6 yields of the control group was 49.8 (5.33-522.46) ng/10 sperm cells. There was no significant difference in median of smallRNA yields of the control group and that of other groups. The median of the other groups with storage temperature0 0 0 6 of 4 C, -20 , and -80 C was 41.09 (7.03-1448.31), 65.95 (7.99-301.16), and 76.42 (10.45-434.25) ng/10 sperm cells,respectively (p-value= 0.314; α=5%). This condition suggested that after purification using SCLB, human sperm cells can be0 0 0 stored at temperatures of 4 C, -20 , or -80 C for seven days, depending on each laboratory facility.