Supplementation of biotin to sperm preparation medium enhances fertilizing ability of spermatozoa and improves preimplantation embryo development
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Motility of spermatozoa helps not only in planning the type of infertility treatment but also directly reflects the success rate in assisted reproductive technology (ART). Previously, biotin, a water-soluble vitamin, has been shown to increase the motility and longevity of cryopreserved human spermatozoa. The present study was designed to understand the molecular basis of the beneficial effects of presence of biotin in sperm wash medium on early embryo development.
The effect biotin supplementation to sperm wash medium on the sperm parameters were assessed in swim-up fraction of normozoospermic and asthenozoospermic ejaculates collected from infertile men. Fertilization and early embryo development was studied using Swiss albino mice.
Even though both biotin and pentoxifylline (PTX) enhanced the motility of spermatozoa from normozoospermic and asthenozoospermic samples, biotin group exhibited higher in vitro survival. Using mouse model, we observed that presence of biotin or PTX in sperm wash medium improved the fertilization rate and blastocyst rate compared to control. Blastocysts from these groups had significantly higher total cell number (P < 0.01) and lower apoptotic index. In silico target prediction revealed that GTPase HRas (HRas), tyrosine-protein phosphatase nonreceptor type 1 (PTP1B), and glucokinase are the probable targets for biotin. Solution-state Nuclear Magnetic Resonance (NMR) studies confirmed that biotin interacts both with human HRas and PTP1B.
Our results indicate that presence of biotin in sperm wash medium can improve the fertilization potential and preimplantation embryo development and can be considered as a safe alternate to PTX.
KeywordsSperm motility Biotin Pentoxifylline Fertilization Blastocyst rate
The financial assistance from Indian Council of Medical Research (ICMR) is thankfully recognized (ICMR/5/10/10/2009-RHN).
Compliance with ethical standards
Study was approved by Institutional Ethics Committee of Kasturba Medical College, Manipal Academy of Higher Education, Manipal (IEC 087/2010), and a written informed consent was taken from subjects willing to provide their semen samples for the study. All the methods were performed in accordance with the institutional guidelines and Helsinki declaration. The study was approved by Institutional Animal Ethics Committee of Kasturba Medical College, Manipal (IAEC/KMC/40/2012). Animal care and handling were conducted according to the institutional guidelines for animal experimentation.
Conflict of interest
The authors declare that they have no competing interests.
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