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

Berliana Hamidah, Ashon Sa'adi, Rina Yudiwati

Abstract


The aim of this study is to learn about the choice of storage temperature for human sperm cells after sperm purification using somatic cell lysis buffer (SCLB) before sperm small ribonucleic acid (RNA) isolation and analysis. This study was true laboratory experiment using the post-test only control group design. The samples were 13 fresh human semen that has been purified using SCLB. The sperm cells then diluted and divided into four aliquots with different treatment. First aliquot that served as a control group was immediately purified while the last three aliquots were stored for 7 days at different temperature: 4oC, -20o, and -80oC. The small RNA yields between each group then compared after the small RNA isolation and measured using microvolume spectrophotometer. The small RNA yields of control group was 49.8 (5.33-522.46) ng/106 sperm cells. It wasn’t show any significant difference with the other groups of storage temperature: 4oC, -20o, and -80oC [41.09 (7.03-1448.31); 65.95 (7.99-301.16); 76.42 (10.45-434.25); p value 0.314] with p value > α (α = 5%). This condition shows that human sperm cells after sperm purification using SCLB can be stored at 4oC, -20o, or -80oC temperatures depends on each laboratory facilities.

Keywords


sperm RNA analysis; purified sperm cells; storage temperature

References


Agarwal A, Mulgund A, Hamada A, Chyatte MR. A unique view on male infertility around the globe. Reprod Biol Endocrinol. 2015;13(37):1–9.

Anton E, Krawetz SA. Spermatozoa as biomarkers for the assessment of human male infertility and genotoxicity. Syst Biol Reprod Med. 2012;58:41–50.

Ramasamy R, Scovell JM, Kovac JR, Cook PJ, Lamb DJ, Lipshultz LI. Fluorescence in situ hybridization detects increased sperm aneuploidy in men with recurrent pregnancy loss. Fertil Steril. 2015;103:906–10.

Agarwal A, Majzoub A, Esteves SC, Ko E, Ramasamy R, Zini A. Clinical utility of sperm DNA fragmentation testing: practice recommendations based on clinical scenarios. Transl Androl Urol. 2016;5(6):935–50.

Jodar M, Sendler E, Moskovtsev SI, Librach CL, Goodrich R, Swanson S, et al. Absence of sperm RNA elements correlates with idiopathic male infertility. Sci Transl Med. 2015;7(295):1–8.

Ellervik C, Vaught J. Preanalytical Variables Affecting the Integrity of Human Biospecimens in Biobanking. Clin Chem. 2015;61(7):914–34.

Mao S, Goodrich RJ, Hauser R, Schrader SM, Chen Z, Krawetz SA. Evaluation of the effectiveness of semen storage and sperm purification methods for spermatozoa transcript profiling. Syst Biol Reprod Med. 2013;59:287–95.

Barragan M, Martınez A, Llonch S, Pujol A, Vernaeve V, Vassena R. Effect of ribonucleic acid (RNA) isolation methods on putative reference genes messenger RNA abundance in human spermatozoa. Andrology. 2015;3:797–804.

Mao S, Sendler E, Goodrich RJ, Hauser R, Krawetz SA. A comparison of sperm RNA-seq methods. Syst Biol Reprod Med. 2014;60(5):308–15.

WHO. WHO laboratory manual for the Examination and processing of human semen. 5 th. Cooper TG, editor. Geneva: WHO; 2010.

Hubel A, Spindler R, Skubitz APN. Storage of human biospecimens: Selection of the optimal storage temperature. Biopreserv Biobank. 2014;12(3):165–75.

Ikeda K, Ichihara K, Hashiguchi T, Hidaka Y, Kang D, Maekawa M, et al. Evaluation of the short-term stability of specimens for clinical laboratory testing. Biopreserv Biobank. 2015;13(2):135–43.

Olivieri EHR, Franco L de A, Pereira RG, Mota LDC, Campos A nio HJFM, Carraro DM. Biobanking practice: RNA storage at low concentration affects integrity. Biopreserv Biobank. 2014;12(1):46–51.




DOI: http://dx.doi.org/10.24293/ijcpml.v26i2.1530

Refbacks

  • There are currently no refbacks.