Method for extracting viable sperms from a semen sample
Inventors
Dendukuri, Dhananjay • Kalme, Satish • Prakash, Vijetha Nagendra • Prasath, Gokul R • Srini, Vasan Satya
Assignees
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Publication Number
US-10465164-B2
Publication Date
2019-11-05
Expiration Date
Abstract
The method provides a hyaluronic-acid binding based technique to separate functionally competent and mature sperm. The method for extracting viable sperms from a semen sample comprises incubating functionalized HA with functionalized paramagnetic beads to obtain a plurality of HA coated paramagnetic beads. The method further comprises incubating the plurality of HA coated paramagnetic beads with the semen sample to obtain a first population and a second population of sperm. The first population comprises sperm bound to the plurality of HA coated paramagnetic beads and the second population comprises sperms not bound to the plurality of HA coated paramagnetic beads. The method comprises magnetically separating the first population from the second population and separating sperms from the plurality of HA coated paramagnetic beads in the separated first population to obtain the viable sperms.
Core Innovation
The invention provides a method for extracting viable sperms from a semen sample using functionalized hyaluronic acid (HA) and functionalized paramagnetic beads. Functionalized HA is incubated with functionalized paramagnetic beads to obtain a plurality of HA coated paramagnetic beads, which bind viable sperm to form a first population and leave sperms not bound as a second population.
After binding, the method magnetically draws the first population away from the second population to separate the second population from the first population and obtain a separated first population. The separated first population comprises viable sperm bound to the plurality of HA coated paramagnetic beads.
The method further enzymatically degrades the HA with a hyaluronidase enzyme to release the sperm from the paramagnetic beads in the separated first population. Finally, the method magnetically draws the paramagnetic beads away from the released sperms to obtain the viable sperms.
Claims Coverage
The document includes one independent claim describing a complete workflow with four inventive features for HA-based viability capture, magnetic separation, hyaluronidase-based release, and magnetic recovery.
Viable sperm capture with HA-coated paramagnetic beads
Incubating functionalized hyaluronic acid (HA) with functionalized paramagnetic beads to obtain a plurality of HA coated paramagnetic beads, and incubating the plurality of HA coated paramagnetic beads with the semen sample to obtain a first population comprising viable sperm bound to the plurality of HA coated paramagnetic beads and a second population comprising sperms not bound to the plurality of HA coated paramagnetic beads.
Magnetically separating bound viable sperms from unbound sperms
Magnetically drawing the first population away from the second population to separate the second population from the first population and to obtain a separated first population, wherein the separated first population comprises viable sperm bound to the plurality of HA coated paramagnetic beads.
Enzymatically degrading HA to release sperm
Enzymatically degrading the HA with hyaluronidase enzyme to release the sperm from the paramagnetic beads in the separated first population.
Magnetically removing paramagnetic beads to obtain viable sperms
Magnetically drawing the paramagnetic beads away from the released sperms to obtain the viable sperms.
Claim 1 covers a combined system in which viable sperm are captured by HA-coated functionalized paramagnetic beads, separated magnetically from unbound sperms, released by hyaluronidase-mediated enzymatic HA degradation, and recovered by a second magnetic draw that removes the paramagnetic beads.
Stated Advantages
Improved DNA fragmentation index (DFI) while maintaining motility and morphology.
Documented Applications
Assisted reproductive technology (ART) applications are described, including IUI, IVF, and ICSI.
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