Development of an adipose-derived stem cell isolation matrix

描述（由申请人提供）：在美国每年大约有50万例植骨手术。自体髂骨移植仍然是金标准，因为它提供了骨形成的基本元素：祖细胞、骨传导基质和骨诱导分子。然而，髂嵴摘除术会带来大量并发症，而且通常提供的移植物体积不足。为了克服这些限制，多能间充质干细胞（MSCs）被添加到骨移植替代品中，并探索在组织工程应用中的应用，如脊柱融合和骨缺损修复。骨髓间充质干细胞（BMSCs）具有向中胚层起源的细胞和组织分化的能力；然而，它们在处理过的骨髓的单个核部分中的频率非常低。脂肪源性干细胞（ASCs）已被提议作为骨髓间充质干细胞的替代品，用于细胞治疗，因为在处理过的抽脂液的基质血管部分（SVF）中发现的多能性ASCs的频率可能比抽髓液的单核部分（BMA）高出数百倍，这取决于供体。目前只有几个基本的护理点设备来集中祖细胞，这是促进骨愈合的基本要素。然而，这些设备存在局限性，因为它们价格昂贵，而且它们要么富集非特异性细胞群，可能会稀释出所需的细胞，要么使用抗体富集特定细胞群，因此存在安全问题。在这个第二阶段的提案中，我们将开发一种用于骨移植的即时自体ASC富集系统，该系统消除了植入前体外细胞扩增的需要。

DESCRIPTION (provided by applicant): Approximately 500,000 bone-grafting procedures are performed each year in the United States. Autologous iliac crest bone graft continues to be the gold standard, because it provides essential elements for bone formation: progenitor cells, an osteoconductive matrix, and osteoinductive molecules. However, iliac crest harvest is associated with a significant number of complications and often provides an inadequate volume of graft. To overcome these limitations, multipotent Mesenchymal Stem Cells (MSCs) have been added to bone graft substitutes and explored for use in tissue engineering applications, such as spinal fusions and bone defect repairs. Bone marrow derived mesenchymal stem cells (BMSCs) are well characterized in their ability to differentiate into cells and tissues of mesodermal origin; however, their frequency in the mononuclear fraction of processed bone marrow is very low. Adipose derived stem cells (ASCs) have been proposed as an alternative to BMSCs for cell therapy because the frequency of multipotent ASCs found in the stromal vascular fraction (SVF) of processed lipoaspirate can be several hundred-fold greater than the mononuclear fraction of bone marrow aspirate (BMA), depending on the donor. Currently there are only a few basic point-of-care devices to concentrate progenitor cells, which are essential elements to foster bone healing. However, these devices are limiting because they are expensive and because they either enrich a non-specific cell population potentially diluting out desired cells or they enrich a specific cell population using antibodies and consequently have safety concerns. In this phase 2 proposal, we will develop a point-of-care autologous ASC enrichment system for bone grafting that eliminates the need for ex vivo cell expansion prior to implantation. PUBLIC HEALTH RELEVANCE: Bone grafting is widely used in orthopaedic surgery to treat fractures and large bone defects and to achieve spinal fusions. However, harvesting a bone graft from patients is associated with a significant number of complications and often provides an inadequate amount of tissue. While alternatives to bone grafts have shown promise, there is still room for considerable improvement. Therefore, a device that could directly isolate progenitor cells from complex biological fluids, and then be used at point-of-care in the operating room, would be of significant clinical benefit.