Supplementation of biotin to sperm preparation medium increases the motility and longevity in cryopreserved human spermatozoa
To study the effect of supplementing biotin to sperm preparation medium on the motility of frozen-thawed spermatozoa.
Semen samples of men attending the University infertility clinic (n = 105) were cryopreserved using glycerol-egg yolk-citrate buffered cryoprotective medium in liquid nitrogen. After a period of two weeks, the semen samples were thawed and the motile spermatozoa were extracted by swim-up technique using Earle’s balanced salt solution (EBSS) medium supplemented with either biotin (10 nM) or pentoxifylline (1 mM). The post-wash motility was observed up to 4 h after incubation.
Both biotin and pentoxifylline supplementation resulted in significant increase in total motility (p < 0.05), progressive motility (p < 0.001) and rapid progressive motility (p < 0.05 v/s biotin and p < 0.01 v/s pentoxifylline) compared to the control at 1 h post-incubation period. Significantly higher percentage of total (p < 0.01, p < 0.05 in biotin and pentoxifylline respectively), progressive (p < 0.001) and rapid progressive motilities (p < 0.01) were observed in these two groups even at 2 h compared to the control. In the control group at 4 h after incubation, ~11% decline in total motility and ~8% decline in progressive motility was observed. However, in both biotin and pentoxifylline group the motility was significantly higher than control (p < 0.001). No significant difference in the motility was observed between biotin and pentoxifylline groups at any of the time intervals studied.
Biotin can enhance the sperm motility and prolong the survival of frozen-thawed semen samples which may have potential benefit in assisted reproductive technology field.
KeywordsBiotin pentoxifylline spermatozoa cryopreservation post-thaw motility
Authors kindly acknowledge the technical support provided by Mrs. Jalyalaxmi Pai, Mrs. Kirthi Patil, and Ms. Anita Poojary. Authors thank Dr. Prashanth Naik and Mr. Naveenchandra Kumar, Mangalore University for editing the language of the manuscript.
- 7.World Health Organization. Laboratory manual for the examination of human semen and sperm-cervical mucus interaction. Cambridge: Cambridge University Press; 1999.Google Scholar
- 10.Mahadevan M, Baker G. Assessment and preparation of semen for in vitro fertilization. In: Wood C, Trounson A, editors. Clinical in vitro fertilization. Berlin: Springer; 83. p. 97–1984.Google Scholar
- 25.Zempleni J, Teixeira DC, Kuroishi T, Cordonier EL, Baier S. Biotin requirements for DNA damage prevention. Mutat Res 2011;doi: mrfmmm2011.08.001/mrfmmm 2011.08.001.