Evaluate a novel product to simultaneously remove dead cells, cellular membranes,

评估一种同时去除死细胞、细胞膜、

• 批准号: 8310392
• 负责人: CHRISTOPHER P REINHARDT
• 金额: $ 14.79万
• 依托单位: BIOPAL, INC
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-09-17 至 2014-03-16
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8310392
• 关键词: Affinity; Antibody Formation; Binding; Buffers; Businesses; Calcium; Cell Culture Techniques; Cell Line; Cell Nucleus; Cell Therapy; Cell membrane; Cells; Cellular Membrane; Collection; Colloids; Diagnostic; Disadvantaged; Effectiveness; Endotoxins; Excision; Industry; Magnetism; Maintenance; Marketing; Membrane; Methods; Nucleic Acids; Oligonucleotides; Performance; Phase; Phosphatidylserines; Phospholipids; Polymers; Preparation; Procedures; Proteins; Reproductive Medicine; Sampling; Serum; Services; Small Business Innovation Research Grant; Stem cells; Sterility; Surface; Technology; Time; Vertebral column; annexin A5; base; clinical application; clinical practice; cost; design; flexibility; inorganic phosphate; magnetic field; nanoparticle; new technology; novel; particle; stem cell therapy

DESCRIPTION (provided by applicant): 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.

描述（由申请人提供）：我们假设可以开发具有商业竞争力的磁性纳米颗粒，以消除细胞培养物（如干细胞）制备中的碎片。目前从健康培养物中去除不需要的物质的方法依赖于使用基于膜联蛋白V的磁性颗粒的磁性负选择。膜联蛋白V与存在于死细胞和细胞膜碎片表面上的磷脂酰丝氨酸（PS）结合。然后施加磁场以从健康培养物中收集死细胞。尽管在商业上是成功的，但基于膜联蛋白V的技术具有显著的缺点：（i）其仅限于除去含PS的膜，（ii）不能除去剥离的核和寡核苷酸，（iii）膜联蛋白V不能高压灭菌，（iv）膜联蛋白V活性可随时间降解，（v）膜联蛋白V需要含钙缓冲液，（vi）难以用于血清，（vii）由于膜联蛋白V的脱落而固有地不稳定，（viii）可能不具有高的内毒素水平，（ix）制造和维护昂贵，因此给最终用户增加了显著的成本。我们提出的产品将消除死亡细胞和细胞膜碎片从文化作为有效的膜联蛋白V为基础的磁性颗粒。我们的产品还将（i）靶向和消除核酸，（ii）在缓冲液选择方面提供更大的灵活性，以及（iii）耐受高压灭菌以确保无菌性。因为在这种情况下， 随着技术成熟进入临床实践，细胞样本制备将变得更加严格，这一应用的成功完成将对生物医学的许多领域产生深远和积极的影响，包括生殖医学，诊断应用和细胞治疗，如自体干细胞治疗。 公共卫生相关性：该项目将评估一种新技术，以从健康细胞培养物中去除不需要的和有害的碎片。由于随着技术成熟进入临床实践，制备细胞样本的条件将变得更加严格，因此我们的产品将对生物医学的许多领域产生深远而积极的影响，包括生殖医学，诊断应用和细胞治疗。