Point of Care Adult Stem Cell Isolation

护理点成人干细胞分离

• 批准号: 7271027
• 负责人: Hanne Gron
• 金额: $ 28万
• 依托单位: AFFINERGY ,INC
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-08-01 至 2009-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7271027
• 关键词: Address; Affinity; Aspirate substance; Autologous; Binding; Biocompatible Materials; Biological; Biological Assay; Biopolymers; Bone Marrow; Bone Marrow Aspiration; Bone Regeneration; Bone Transplantation; Cartilage; Cell Count; Cell Separation; Cell Survival; Cell Therapy; Cell physiology; Cells; Chemistry; Clinical; Development; Device Designs; Devices; Elements; Enzymes; Fatty acid glycerol esters; Filtration; Fluorescence; Fostering; Generations; Goals; Gold; Growth Factor; Harvest; Healed; Hour; Human; Label; Marrow; Mediating; Mesenchymal Stem Cells; Metals; Methods; Muscle; Numbers; Organ Transplantation; Osteogenesis; Peptides; Phage Display; Phase; Plastics; Polymers; Polystyrenes; Population; Process; Range; Research; Site; Small Business Funding Mechanisms; Small Business Innovation Research Grant; Specific qualifier value; Specificity; Standards of Weights and Measures; Stem cells; Surface; Suspension substance; Suspensions; System; Technology; Tendon structure; Testing; Therapeutic; Thrombin; Tissues; To specify; United States; Work; adult stem cell; base; bone; bone healing; bone morphogenic protein; cell growth; chemical release; crosslink; design; healing; implantation; improved; interfacial; novel; osteoinductive factor; point of care; protein aminoacid sequence; soft tissue

DESCRIPTION (provided by applicant): Approximately 500,000 bone-grafting procedures are performed each year in the United States (Boden, 2002). Autologous iliac crest bone graft continues to be the gold standard because it provides the three essential elements for bone formation: osteoprogenitor cells, an osteoconductive matrix, and osteoinductive molecules. Iliac crest harvest is however associated with a significant number of complications and often provides an inadequate volume of graft (Gupta 2001). To address these limitations, much work has been done to improve alternative grafts with better osteoconductive matrices and exogenous delivery of osteoinductive agents such as the bone morphogenic proteins (BMP's) (Sykaras and Opperman 2003). In contrast, there are only a few basic point-of-care devices to isolate osteogenic progenitor cells, which are the third and truly essential element to foster bone healing. A device that could isolate stem cells from a bone marrow aspirate (BMA) at point of care would be of significant clinical benefit. In this phase I proposal, we will identify specific binding peptides for the multipotent subpopulation of cells in human bone marrow known as mesenchymal stem cells (MSC's), which have been shown to promote healing of bone and soft tissue (Caplan and Bruder 2001). Interfacial biomaterials (IFBM's) will be synthesized by chemically crosslinking combinations of MSC binding sequences with Affinergy's polystyrene binding sequences. These IFBM's will then be tested for their ability to bind and retain MSC's from suspensions of MSC's, suspensions of mixed cell populations, and human bone marrow aspirates. Finally, we will develop a chemical release system for bound MSC's and verify their continued viability and multipotentiality after release. These studies, upon successful completion, will validate the utility of IFBM technology in the development of a point of care stem cell isolation device. Although beyond the scope of this proposal, we envision the addition of IFBM's that bind osteogenic growth factors, which are also present in bone marrow (Barnes, Kostenuik et al. 1999), to a second generation bone marrow filtration device. This second generation device is envisioned to select and concentrate osteoinductive molecules as well as a functional population of osteoprogenitor cells. Finally, to be most effective as a timely point of care device, the entire process from drawing a marrow aspirate to re-implantation of the filtered osteogenic cells and growth factors should be as quick as possible (ideally, less than 1 hour).

描述（由申请人提供）：在美国，每年进行大约500,000个骨夹程序（Boden，2002）。自体峰骨移植物仍然是黄金标准，因为它为骨形成提供了三个基本要素：骨源细胞，骨导导基质基质和骨诱导分子。然而，iLiac Crest收获与大量并发症有关，并且通常提供的移植物数量不足（Gupta 2001）。为了解决这些局限性，已经进行了许多工作来改善替代移植物，并具有更好的骨导导式基质以及骨诱导剂（例如骨形态发生蛋白（BMP）（Sykaras and Opperman 2003））的骨诱导剂的外源递送。相比之下，只有少数基本的护理设备可以分离成骨的祖细胞，这是促进骨骼愈合的第三个且真正必不可少的元素。可以在护理点上分离出干细胞（BMA）的干细胞的装置将具有重要的临床益处。在此阶段I建议中，我们将确定特定的结合肽，用于人类骨髓中细胞的多能亚群（称为间充质干细胞）（MSC）（MSC），这些肽已被证明可以促进骨骼和软组织的愈合（Caplan和Bruder 2001）。界面生物材料（IFBM）将通过使用Affinergy的聚苯乙烯结合序列的MSC结合序列的化学交联组合来合成。然后，将对这些IFBM的结合和保留MSC从MSC的悬浮液，混合细胞种群的悬浮液以及人骨髓抽吸的能力进行测试。最后，我们将开发一个用于绑定MSC的化学释放系统，并验证其释放后的持续生存力和多能性。这些研究成功完成后将验证IFBM技术在开发干细胞隔离设备的开发中的实用性。尽管除了该提案的范围之外，我们还设想添加结合成骨生长因子的IFBM，这些因子也存在于骨髓中（Barnes，Kostenuik等，1999），以及第二代骨髓过滤器。设想这款第二代装置可以选择和浓缩骨诱导分子以及骨基源细胞的功能群体。最后，要成为及时的护理设备，最有效的是，从骨髓抽吸到重新植入过滤的成骨细胞和生长因子的整个过程应尽快（理想情况下，少于1小时）。