Development of an adipose-derived stem cell isolation matrix

描述（由申请人提供）：在美国，每年大约进行500，000例骨移植手术。自体髂骨移植仍然是金标准，因为它提供了骨形成的基本要素：祖细胞，骨传导基质和骨诱导分子。然而，髂嵴采集与大量并发症相关，并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（MSC）已被添加到骨移植替代品中，并探索用于组织工程应用，如脊柱融合和骨缺损修复。骨髓源性间充质干细胞（BMSC）的特征在于其分化成中胚层来源的细胞和组织的能力;然而，它们在经处理的骨髓的单核部分中的频率非常低。脂肪源性干细胞（ASC）已被提出作为BMSC的替代品用于细胞治疗，因为在经处理的脂肪抽吸物的基质血管部分（SVF）中发现的多能ASC的频率可以是骨髓抽吸物（BMA）的单核部分的几百倍，这取决于供体。目前，只有几种基本的定点护理设备可以集中祖细胞，这是促进骨愈合的基本要素。然而，这些装置是有限制的，因为它们是昂贵的，并且因为它们或者富集非特异性细胞群体，潜在地稀释所需细胞，或者它们使用抗体富集特异性细胞群体，因此具有安全性问题。在这个2期提案中，我们将开发一种用于骨移植的即时自体ASC富集系统，该系统消除了在植入前进行体外细胞扩增的需要。 公共卫生相关性：骨移植广泛用于骨科手术，以治疗骨折和大面积骨缺损，并实现脊柱融合。然而，从患者身上获取骨移植物与大量并发症相关，并且通常提供的组织量不足。虽然骨移植的替代品已显示出希望，但仍有相当大的改进空间。因此，一种可以直接从复杂的生物液体中分离祖细胞，然后在手术室的护理点使用的设备将具有显著的临床益处。

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.