Selective enrichment of high-quality DNA intact sperms from a semen sample
Inventors
Soman, Raunak Jay • Soman, Jay • Mahesh, Motukuri
Assignees
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Publication Number
US-11926847-B2
Publication Date
2024-03-12
Expiration Date
Abstract
Disclosed is a composition for selective enrichment of progressively motile non-fragmented/intact DNA sperms during swim-up of semen sample comprising combination of electrolytes, a mix of excipients, polyyvinylpyrrolidone, recombinant human serum albumin or bovine serum albumin. A method of selective enrichment of DNA-intact sperms using the compositions of the present invention is also disclosed. The DNA fragmentation index of the swim-up sperms are less than the DNA fragmentation index of the neat samples. The media compositions of the present invention are applicable for enrichment of non-fragmented/intact DNA sperms for therapeutic purposes in the field of assisted reproductive technologies.
Core Innovation
The disclosed invention relates to a swim-up medium for selective enrichment of fractions containing DNA-intact sperm cells after swim-up, where the DNA fragmentation index of the post swim-up fraction is lower than the DNA fragmentation index of neat semen. The medium is presented as a solvent and a solute composition, with the solvent defined as water and the solute composition including a concentration of up to 15% w/v.
The solute composition includes electrolytes, excipients, Polyvinylpyrrolidone (PVP), and recombinant Human Serum Albumin (rHSA) or bovine serum albumin (BSA). The targeted enriched fraction is described as progressive, DNA-intact, and non-fragmented, and performance is evaluated using a DFI scoring basis linked to SPERM CHROMATIN DISPERSION (SCD) and Halo assessment.
Representative formulations include SNZ-Plain and SNZ-Plus swim-up media, where SNZ-Plus includes PVP and recombinant human serum albumin (rHSA) or bovine serum albumin (BSA) in the solute composition. The description further reports that the medium is configured with a pH range of 7.2–7.6 and that comparative outcomes show lower DNA fragmentation index values than neat semen and outperform a commercial medium.
Claims Coverage
The independent claim covers a swim-up medium for selective enrichment of fractions containing DNA-intact sperm cells post swim-up, requiring that the DFI_post swim-up is lower than the DFI of neat semen. The main inventive features are defined by the specific solute, electrolyte, excipient, PVP, and albumin ingredient classes and concentration limits, with an explicit water solvent.
Swim-up medium with selective DNA-intact enrichment based on DFI
A swim-up medium for selective enrichment of fractions containing DNA-intact sperm cells post swim-up, the fraction having a DNA fragmentation index where DFI_post swim-up < DFI_neat, from a semen sample, comprising a solvent and a solute composition; the solvent is water.
Solute composition up to 15% w/v with defined ingredient classes
The solute composition is at a concentration of up to 15% w/v and comprises electrolytes, excipients, Polyvinylpyrrolidone (PVP), and recombinant Human Serum Albumin (rHSA) or bovine serum albumin (BSA) within specified concentration ranges.
Electrolyte combination within specified concentration range
A combination of electrolytes at a concentration range of 10% to 50% w/w consisting of sodium chloride, potassium chloride, magnesium sulphate, potassium di-hydrogen orthophosphate, calcium chloride, sodium bicarbonate, sodium phosphate monobasic anhydrous, magnesium chloride and glucose.
Excipients combination within specified concentration range
A combination of excipients at a concentration range of 1% to 30% w/w consisting of phenol red, HEPES buffer, gentamicin, sodium pyruvate, sodium lactate, glutamine, glycine, L-Arginine, and methionine.
PVP concentration range
Polyvinylpyrrolidone (PVP) at a concentration range of 3% to 80% w/w.
rHSA or BSA concentration range
Recombinant Human Serum Albumin (rHSA) or bovine serum albumin (BSA) at a concentration range of 0.1% to 50% w/w.
Across the independent claim, the inventive coverage centers on a water-solvent swim-up medium with a solute composition up to 15% w/v defined by electrolyte and excipient combinations plus PVP and rHSA or BSA, and with a required DNA fragmentation index outcome criterion for the post swim-up fraction.
Stated Advantages
Selectively enriches fractions containing DNA-intact sperm cells post swim-up with a lower DNA fragmentation index than neat semen.
Described comparative performance: SNZ-Plus decreases DFI relative to neat semen and outperforms a commercial medium.
Documented Applications
Assisted reproductive technologies (ART), including IVF, ICSI, IMSI, GIFT, ZIFT, Gamete intra fallopian transfer, and Zygote intra fallopian transfer.
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