Point of Care Adult Stem Cell Isolation

描述（由申请人提供）：美国每年进行约500，000例骨移植手术。5自体髂嵴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。6为了解决这些局限性，已经做了大量工作来改进替代移植物，其具有更好的骨传导基质和骨诱导剂（如骨形态发生蛋白（BMP））的外源性递送。7目前只有少数基本的即时装置来分离祖细胞，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。因此，一种安全、廉价的装置，可以在床旁从骨髓穿刺液（BMA）或其他组织基质中富集成体干细胞，将具有显著的临床益处。 在第二阶段的建议中，我们将继续开发一种细胞分离装置，以丰富人类骨髓中称为间充质干细胞（MSC）的多能细胞亚群。（*2）此外，我们将通过展示我们可以从脂肪组织中捕获成体干细胞来证明分离装置的多功能性。脂肪组织的优点是可获得更多数量的骨愈合细胞。我们已经鉴定并合成了一种MSC结合肽，可以从骨髓和脂肪组织中分离成体干细胞，这些分离的细胞是多能的。MSC结合肽将共价连接到固体支持基质上，并将形成细胞分离装置的功能核心。将使用原型装置捕获和释放分离的细胞，并在大鼠颅骨缺损的骨愈合中测试不同浓度的细胞。分离的MSC库具有在一系列适应症中发挥关键临床功能的潜力，所述适应症包括骨关节炎、Chron病和软组织修复，例如肌腱、韧带或伤口。该产品将为自体细胞的制备提供一种安全且具有成本效益的床旁隔离系统。

DESCRIPTION (provided by applicant): Approximately 500,000 bone-grafting procedures are performed each year in the United States.5 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.6 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 morphogenetic proteins (BMPs).7 Currently there are only a few basic point-of-care devices to isolate 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. Therefore, a safe, inexpensive device that could enrich adult stem cells from a bone marrow aspirate (BMA) or other tissue stroma at point-of-care would be of significant clinical benefit. In this phase II proposal, we will continue to develop a cell isolation device to enrich the multipotent subpopulation of cells in human bone marrow known as mesenchymal stem cells (MSCs). (*2)Furthermore, we will demonstrate versatility of the isolation device by showing that we can capture adult stem cells from adipose tissue. The advantage of adipose tissue is the availability of greater numbers of bone healing cells. We have identified and synthesized an MSC-binding peptide that can isolate adult stem cells from bone marrow and adipose tissue, and these isolated cells are multipotent. The MSC-binding peptide will be covalently attached to a solid support matrix and will form the functional core of the cell isolation device. The cells isolated will be captured and released using the prototype device, and different concentrations of cells will be tested in bone healing of a rat calvarial defect. An isolated pool of MSCs has the potential to serve key clinical functions in a range of indications including osteoarthritis, Chron's disease and soft tissue repair such as tendon, ligament or wounds. This product will provide a safe and cost-effective point-of-care isolation system for the preparation of autologous cells.