We hypothesize that a commercially competitive magnetic nanoparticle can be developed to eliminate debris in the preparation of cell cultures, such as stem cells. Current methods to remove unwanted material from healthy cultures rely on magnetic negative selection using Annexin V-based magnetic particles. Annexin V binds to the phospholipid phosphatidylserine (PS) present on the surface of dead cells and cell membrane fragments. A magnetic field is then applied to collect dead cells from the healthy culture. Although commercially successful, Annexin V-based technology has significant drawbacks: (i) it is restricted to removing only PS-containing membranes, (ii) stripped nuclei and oligonucleotides cannot be removed, (iii) Annexin V cannot be autoclaved, (iv) Annexin V activity can degrade over time, (v) Annexin V requires calcium-containing buffers, (vi) is difficul to use in serum, (vii) is inherently unstable due to the sloughing of Annexin V, (viii) is likely t have high endotoxin levels, (ix) is expensive to manufacture and maintain, therefore adding significant cost to the end- user. Our proposed product will eliminate dead cells and cell membrane fragments from culture as effectively as Annexin V-based magnetic particles. Our product will also (i) target and eliminate nucleic acids, (ii) provide greater flexibility in buffe selection, and (iii) tolerate autoclaving to assure sterility. Because the conditions under which a cell sample is prepared will become more stringent as technologies mature into clinical practice, successful completion of this application will have a profound and positive impact on many sectors of biomedicine, including reproductive medicine, diagnostic applications and cell therapy, such as autologus stem cell therapy.
Public Health Relevance: The project will evaluate a novel technology to remove unwanted and harmful debris from healthy cell cultures. Because the conditions under which a cell sample is prepared will become more stringent as technologies mature into clinical practice, our product will have a profound and positive impact on many sectors of biomedicine, including reproductive medicine, diagnostic applications and cell therapy.
Public Health Relevance Statement: The project will evaluate a novel technology to remove unwanted and harmful debris from healthy cell cultures. Because the conditions under which a cell sample is prepared will become more stringent as technologies mature into clinical practice, our product will have a profound and positive impact on many sectors of biomedicine, including reproductive medicine, diagnostic applications and cell therapy.
NIH Spending Category: Bioengineering; Biotechnology; Nanotechnology
Project Terms: Affinity; annexin A5; Antibody Formation; base; Binding (Molecular Function); Buffers; Businesses; Calcium; Cell Culture Techniques; Cell Line; Cell membrane; Cell Nucleus; Cell Therapy; Cells; Cellular Membrane; clinical application; clinical practice; Collection; Colloids; cost; design; Diagnostic; Disadvantaged; Effectiveness; Endotoxins; Excision; flexibility; Industry; inorganic phosphate; magnetic field; Magnetism; Maintenance; Marketing; Membrane; Methods; nanoparticle; new technology; novel; Nucleic Acids; Oligonucleotides; particle; Performance; Phase; Phosphatidylserines; Phospholipids; Polymers; Preparation; Procedures; Proteins; Reproductive Medicine; Sampling; Serum; Services; Small Business Innovation Research Grant; stem cell therapy; Stem cells; Sterility; Surface; Technology; Time; Vertebral column
