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。众所周知，骨髓中的细胞具有骨再生能力，但它们的数量有限。通过使用体外扩增技术增加这些骨祖细胞的数量，并将这些细胞与基质材料结合起来，可能会显著增加新骨形成的数量和速度，并取代自体移植的需要。第一阶段研究确定了骨祖细胞扩增的扩张变量，并表明这一过程可以扩大到自动化的临床规模细胞培养系统--AstromReplicell(R)细胞生产系统(ARS)。在第二阶段研究的目标1中，将继续优化骨祖细胞的扩张过程。优化将不仅通过体外和体内的生物学性能来衡量，而且还将通过扩大过程的商业化程度来衡量。具体地说，将确定其他细胞群体在骨祖细胞扩增和功能中的作用，将确定在培养中包括外源因素的最低要求，将确定支持从小容量骨髓抽提物扩增的接种密度和最大喂养计划，将检查在无动物血清环境中的扩增潜力。在目标2中，这些相同的细胞产品的血管生成潜力将通过集落试验和管形成试验在体外进行测量，因为血管形成对于最佳的骨再生非常重要。目的3将在异位小鼠模型中验证ARS的最佳扩张过程，并评估体内骨和血管的再生。AIM 4将包括一项L/11期临床试验，使用选定的细胞产品与基质材料结合治疗胫骨不愈合骨折。这些研究的成功完成将导致开发一种可用于骨再生的细胞治疗试剂盒。

DESCRIPTION (provided by applicant): Due to morbidity and limited amounts of material, alternatives to autologous grafts for bone repair are desirable. Cells within the bone marrow are known to possess bone regenerative capacity, but they are limited in number. Increasing these osteoprogenitor cell populations through the use of ex vivo expansion technologies and combining these cells with a matrix material may significantly increase the amount and rate of new bone formation, and replace the need for autologous grafts. Phase I studies identified expansion variables for osteoprogenitor cell expansion and showed that this process can be scaled up into the automated clinical-scale cell culture system, the AastromReplicell(r) Cell Production System (ARS). In Aim 1 of the Phase II studies, optimization of the osteoprogenitor expansion process will continue. Optimization will be measured by not only biological performance in vitro and in vivo, but also the ease of commercialization of the expansion process. Specifically, the role of other cell populations in osteoprogenitor cell expansion and function will be determined, the minimal requirements for including exogenous factors in cultures will be identified, inoculation densities and maximized feeding schedules will be determined to support expansion from small volume bone marrow aspirates, the potential for expansion in an animal serum-free environment will be examined. In Aim 2, the angiogenic potential of these same cell products will be measured in vitro by colony assay and tube formation assays, as blood vessel formation is important for optimal bone regeneration. Aim 3 will validate optimal expansion processes in the ARS and evaluate bone and vascular regeneration in vivo in an ectopic mouse model. Aim 4 will include a Phase l/ll clinical trial using the selected cell product in combination with matrix material to treat tibial non-union fractures. Successful completion of these studies will lead to the development of a cell therapy kit that can be used for bone regeneration.