Sperm selection is an essential component of all assisted reproductive treatments(ARTs)and is by far the most neglected step in the ART workflow in regard to technological innovation.Conventional sperm selection metho...Sperm selection is an essential component of all assisted reproductive treatments(ARTs)and is by far the most neglected step in the ART workflow in regard to technological innovation.Conventional sperm selection methodologies typically produce a higher total number of sperm with variable motilities,morphologies,and levels of DNA integrity.Gold-standard techniques,including density gradient centrifugation(DGC)and swim-up(SU),have been shown to induce DNA fragmentation through introducing reactive oxygen species(ROS)during centrifugation.Here,we demonstrate a 3D printed,biologically inspired microfluidic sperm selection device(MSSP)that utilizes multiple methods to simulate a sperms journey toward selection.Sperm are first selected based on their motility and boundary-following behavior and then on their expression of apoptotic markers,yielding over 68%more motile sperm than that of previously reported methods with a lower incidence of DNA fragmentation and apoptosis.Sperm from the MsSP also demonstrated higher motile sperm recovery after cryopreservation than that of sU or neat semen.Experiments were conducted side-by-side against conventional SU methods using human semen(n=33)and showed over an 85%improvement in DNA integrity with an average 90%reduction in sperm apoptosis.These results that the platform is easy-to-use for sperm selection and mimics the biological function of the female reproductive tract during conception.展开更多
基金support of the Australian Research Council through Discovery Project Grants(DP200101860)the National Health and Medical Research Council through the Career Development Fellowship(APP1143377)+1 种基金R.N.acknowledges support from the Australian Research Council Discovery Program(DP190100343)Monash Interdisciplinary Research Program.We would like to acknowledge Dr.Sajad Razavi Bazaz for his helpful contribution and assistance in the initial stages of this project.
文摘Sperm selection is an essential component of all assisted reproductive treatments(ARTs)and is by far the most neglected step in the ART workflow in regard to technological innovation.Conventional sperm selection methodologies typically produce a higher total number of sperm with variable motilities,morphologies,and levels of DNA integrity.Gold-standard techniques,including density gradient centrifugation(DGC)and swim-up(SU),have been shown to induce DNA fragmentation through introducing reactive oxygen species(ROS)during centrifugation.Here,we demonstrate a 3D printed,biologically inspired microfluidic sperm selection device(MSSP)that utilizes multiple methods to simulate a sperms journey toward selection.Sperm are first selected based on their motility and boundary-following behavior and then on their expression of apoptotic markers,yielding over 68%more motile sperm than that of previously reported methods with a lower incidence of DNA fragmentation and apoptosis.Sperm from the MsSP also demonstrated higher motile sperm recovery after cryopreservation than that of sU or neat semen.Experiments were conducted side-by-side against conventional SU methods using human semen(n=33)and showed over an 85%improvement in DNA integrity with an average 90%reduction in sperm apoptosis.These results that the platform is easy-to-use for sperm selection and mimics the biological function of the female reproductive tract during conception.