We propose to develop an automated microfluidic microchip merged with a high-throughput cell- encapsulating droplet ejection system for efficient, rapid, and inexpensive blood cryopreservation. Blood is the single most important tissue for biopreservation. In particular, red blood cells (RBCs) are required for transfusion, whenever patients suffer massive blood loss due to: (1) trauma;(2) bleeding disorders;(3) major surgery;or (4) post-partum hemorrhage. Current technology only allows frozen storage of blood and blood products, including packed RBCs. Current blood-freezing technologies employ labor and time intensive procedures that require trained clinical technicians. The complex manual handling involved in blood biopreservation results in high cost, long-processing times, and process variability. Currently, cryopreservation of blood cells is mostly done by slow-freezing. However, slow freezing leaves blood cells susceptible to intracellular damage from intracellular ice crystal formation (IIF). Therefore there is a significant need for improved technologies enabling effective cryopreservation of blood. Although it is shown in the literature that vitrification techniques could achieve better biopreservation outcomes for various cell types such as RBCs and oocytes, vitrification is not applied to blood biopreservation clinically due to throughput limitations. The current vitrification methods require microliter volumes of cells to be filled into straws that are then vitrified, which is not feasible to biopreserve liters of blood. Since these products have limited shelf lives, blood freezing must be done continually and routinely in every hospital and medical center in the US to meet the constant demand. Accordingly, there is a need for a new platform technology that will transform the operational logistics for an efficient future for the existing blood supply chain mechanisms. In this project, we will advance the clinical practice in blood cryopreservation by leveraging the advantages provided by vitrification and enabled by our novel microscale technologies. As a result we expect to achieve: ultra-rapid cooling rates (10,000 oC/sec) at low cryoprotectant agent concentrations with low levels of ice formation. These conditions will lead to improved functionality and longer shelf life of RBCs (>42 days). We are proposing to develop an enabling platform applicable to practically all cell types, especially therapeutic RBCs, peripheral blood stem cells, and primary hepatocytes. If successful, the proposed research can have a significant impact on the long-term storage of blood products for both civilian and military needs.
We apply nano- and micro-scale technologies to develop a novel closed system for automated blood cryopreservation. Such a technology would have a significant clinical impact on blood banking, storage and transportation.
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