A surface-engineered microfluidic device for antibody-mediated negative selection of high-quality sperm for assisted reproduction

Preparation of spermatozoa with optimal developmental competence remains a challenge in assisted reproduction. Conventional techniques based on sperm motility and morphology fail to adequately remove sperm with DNA damage. Here, we report development of a microfluidic device with a functionalized surface, inspired by the physiological processes of immune cell-mediated sperm selection in the female reproductive tract. A plasma-polymerized polyoxazoline (PPOx) film is applied to glass channel slides by deposition of 2-methyl-2-oxazoline, to establish a stable, biocompatible interface confirmed by X-ray Photoelectron Spectroscopy (XPS), ellipsometry, and sperm culture assays. To selectively eliminate pre-apoptotic and apoptotic spermatozoa wherein DNA damage is common, anti-phosphatidylserine (Anti-PS) antibody is immobilized to the PPOx-coated surface proximal to the channel slide inlet, while the sperm chemoattractant progesterone is adsorbed near the outlet. To optimise selective functionality, the surface topography is tailored by covalent immobilization of gold nanoparticles and addition of microchannels. Sperm recovered after processing whole liquified semen then consistently exhibit high motility and morphology, with <1% showing apoptosis-associated membrane damage or DNA fragmentation. Compared with conventional swim-up or other microfluidic approaches, the device yields sperm with improved quality, offering a simple one-step sperm selection strategy with potential for application in human and animal assisted reproduction. Short text and graphic for 45 the Table of Contents (ToC)This study reports a microfluidic device with a functionalized surface utilizing a polyoxazoline coating and covalently immobilized gold nanoparticles and anti-phosphatidylserine antibody. The device selectively eliminates pre-apoptotic and apoptotic spermatozoa and yields sperm with substantially improved quality and low DNA damage, offering a simple one-step sperm selection device with potential for application in human and animal assisted reproduction. O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=166 SRC="FIGDIR/small/673619v1_ufig1.gif" ALT="Figure 1"> View larger version (57K): org.highwire.dtl.DTLVardef@b07026org.highwire.dtl.DTLVardef@192e5b5org.highwire.dtl.DTLVardef@127095aorg.highwire.dtl.DTLVardef@1d70ceb_HPS_FORMAT_FIGEXP M_FIG C_FIG