Ravata Solutions, Inc. SBIR Phase I Award, July 2021

AbstractThe biomedical research community extensively relies on the availability and continued utilization of the rodent research model. With the discovery of CRISPR, and the unprecedented degree with which it allowed scientists to manipulate the genome, an explosion of clinically relevant models has made their way into existence. This leading edge of precision medicine is responsible for the phenotyping of harmful SNP mutations, modeling of disease variants in ethnically distinct populations, and understanding of orphan diseases. Production of these models heavily relies upon in-vitro assisted reproductive techniques including cryo- preservation, gene editing, and in-vitro fertilization however these techniques also reduce the reproductive viability of embryos and the percentage of live-born. As the number of unique models continues to increase facilities require automated and standardized in-vitro assisted reproductive tools to flexibly scale with demand and maintain larger litters. To date embryo handling has been done via mouth pipetting, and embryo viability has been assessed through attrition by either transferring all the available embryos as soon as possible or waiting several days for the damaged cells to eventually arrest leaving some competent blastocysts. Recent developments have measured oocyte/embryo mechanical properties as a non-invasive bio- marker of viability by optically measuring embryo deformation when aspirating through a micro-pipette; however, this technique is still manually demanding and measurement collection systems are imperfect. We propose building a high-throughput microfluidic lab on chip that can identify the viability of embryos immediately after exposure to environmental stresses brought on from cryo-preservation, gene editing, and in-vitro fertilization. Building on the known predictive power of zygote mechanical properties and Ravata’s micro-electrode platform, we would validate an electrical sensor array to electrically quantify embryo displacement through a microfluidic aspirator to measure nuclear and cytoplasmic maturation as an early embryo viability detection system.NarrativeThere is increasing demand on the availability of genetically engineered mouse models to understand and develop treatments to human disease. Unfortunately the time and work put into producing these mice is suboptimal meaning only well funded labs can afford the most up to date and most clinically relevant mice. By developing lab on chip technologies for assisted reproduction we can reduce the cost of working with these research models improving the quality of research from other NIH funded grants.