Clinical-scale Production of Osteoprogenitor Cells

描述（由申请人提供）：由于发病率和材料量有限，需要自体移植物的替代品进行骨修复。骨髓基质细胞（MSC）和载体基质的混合物是已知的，以增强骨修复。通过体外扩增增加MSC的数量可以显著增加新骨形成的量和速率。在本提案中，将测试自动化临床规模细胞培养系统AastromReplicell（tm）细胞生产系统（AR/CPS）产生大量具有成骨潜能的MSC的能力。将通过流式细胞术（STRO-1 +、CD 63+）、体外形成矿化基质的能力和在SCID/NOD小鼠中形成骨的能力鉴定骨祖细胞。使用小规模的培养，参数包括介质灌注率，培养持续时间，和添加剂的影响，如地塞米松，抗坏血酸，FGF-2和BMP-2，将进行研究。然后将在AR/CPS中测试骨祖细胞生产的最佳条件，以确认临床规模的可行性。随着这些研究的成功完成，AR/CPS中产生的MSC将在临床试验中进行测试，以确定这些细胞治疗骨折不愈合的安全性和有效性。此外，这项工作可能会导致新的治疗骨骼疾病，如骨质疏松症和骨关节炎。

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.