Clinical-scale Production of Osteoprogenitor Cells

描述(由申请人提供)：由于发病率和材料的数量有限，替代自体移植物进行骨修复是可取的。已知骨髓基质细胞(MSC)和载体基质的混合物可以增强骨修复。通过体外扩增的方法增加MSC的数量，可能会显著增加新骨形成的数量和速度。在这项提案中，将测试自动化临床规模细胞培养系统AstromReplicell(Tm)细胞生产系统(AR/CPS)产生大量具有成骨潜力的MSC的能力。通过流式细胞仪(Stro-1，CD63)、体外矿化基质形成能力和SCID/NOD小鼠成骨能力鉴定骨祖细胞。使用小规模培养，将研究包括介质灌注率、培养时间以及地塞米松、抗坏血酸、成纤维细胞生长因子-2和骨形态发生蛋白-2等添加剂的影响。然后将在AR/CPS中测试找到的最适合生产骨祖细胞的条件，以确认在临床规模上的可行性。随着这些研究的成功完成，在AR/CPS中产生的MSC将在临床试验中进行测试，以确定这些细胞在治疗骨不连骨折中的安全性和有效性。此外，这项工作可能会导致骨质疏松症和骨关节炎等骨骼疾病的新疗法。

DESCRIPTION (provided by applicant): Due to morbidity and limited amounts of material, alternatives to autologous grafts for bone repair are desirable. Mixtures of marrow stromal cells (MSC) and carrier matrices are known to augment bone repair. Increasing the number of MSC through ex vivo expansion may significantly increase the amount and rate of new bone formation. In this proposal, the ability of an automated clinical-scale cell culture system, the AastromReplicell (tm) Cell Production System (AR/CPS), to generate large numbers of MSC with osteogenic potential will be tested. Osteoprogenitor cells will be identified by flow cytometry (STRO-1 +, CD63+), ability to form mineralized matrix in vitro, and ability to form bone in SCID/NOD mice. Using small-scale cultures, parameters including medium perfusion rates, culture duration, and the effects of additives such as dexamethasone, ascorbate, FGF-2 and BMP-2, will be studied. Conditions found optimal for osteoprogenitor cell production will then be tested in the AR/CPS to confirm feasibility at the clinical scale. With successful completion of these studies, MSC generated in the AR/CPS will be tested in clinical trials to establish the safety and efficacy of these cells in treating nonunion fractures. In addition, this work may lead to novel treatments for skeletal diseases such as osteoporosis and osteoarthritis.