Alveolar Bone Marrow Derived Stem Cells for Clinical Cell Therapy

描述（由申请人提供）：细胞疗法在颅面缺损和畸形的重建和再生方面具有巨大的潜力。细胞治疗的最重要方面之一是鉴定用于分离细胞的最合适的细胞来源。从历史上看，来自骨髓的间充质干细胞（MSC）已在许多临床前研究中显示出再生潜力。然而，用于分离MSC的传统技术通常涉及从髂嵴收获骨髓。由于髂嵴骨髓采集程序的侵入性和该组织的发育（软骨内）起源，更希望有一种更容易获得和发育相似（膜内）的MSC来源用于颅面治疗适应症。为此，我们有初步的证据表明，MSC可以预测分离从牙槽骨骨髓，使用一个简单的，标准化的，可重复的技术。我们的假设是，使用目前的良好生产规范（cGMP）的指导方针，牙槽骨髓源性间充质干细胞（aBMSCs）可以安全地扩大离体，数量足以自体临床移植在人类颅面骨缺损。此外，我们假设，在体外和临床前的替代标志物存在，可以预测临床再生的结果。为了验证这些假设，拟议的项目有三个具体目标（SA）。在SA 1中，我们将从30个不同的人类受试者中分离、扩增和表型表征肺泡骨髓源性干细胞群体。SA 2将使用异位和临床相关原位模型系统评价体内aBMSC的临床前骨再生潜力。最后，在SA 3中，将aBMSC自体移植到人颅面骨缺损中后，我们将确定不同aBMSC群体的表型特征如何与其各自在体内和临床上再生骨的能力相关。主要成果措施包括：表型表征; aBMSC再生骨和血管组织的体内潜力; aBMSC细胞分离和扩增方法用于人应用的安全性; aBMSC再生骨和血管组织的临床潜力;以及体外表型、体内骨再生和临床骨再生之间的相关性检查。这项研究的发现可以显着推进颅面再生的临床细胞治疗方法。此外，使用所采用的技术分离aBMSC将在提供用于干细胞和骨生物学的基础研究的MSC的替代来源方面具有广泛的科学影响。

DESCRIPTION (provided by applicant): Cell therapy holds significant potential for the reconstruction and regeneration of craniofacial defects and deformities. One of the most important aspects of cell therapy is the identification of the most appropriate cell source for isolation of cells. Historically, mesenchymal stem cells (MSCs) derived from bone marrow have demonstrated regenerative potential in many preclinical studies. However, traditional techniques for isolating MSCs typically involve a bone marrow harvest from the iliac crest. Due to the invasive nature of the iliac crest bone marrow harvest procedure and the developmental (endochondral) origin of this tissue, it would be far more desirable to have a more readily accessible and developmentally similar (intramembranous) source of MSCs for craniofacial therapeutic indications. To this end, we have preliminary evidence which suggests that MSCs can be predictably isolated from alveolar bone marrow, using a simple, standardized, reproducible technique. Our hypothesis is that using current Good Manufacturing Practices (cGMP) guidelines, alveolar bone marrow derived MSCs (aBMSCs) can be safely expanded ex vivo, to numbers sufficient for autogenous clinical transplantation in human craniofacial bone defects. Additionally, we hypothesize that in vitro and preclinical surrogate markers exist which can be predictive of clinical regenerative outcomes. To test these hypotheses, the project proposed has three Specific Aims (SAs). In SA1, we will isolate, expand, and phenotypically characterize populations of alveolar bone marrow derived stem cells from 30 different human subjects. SA2 will evaluate the preclinical bone regenerative potential of aBMSCs in vivo using ectopic and clinically relevant orthotopic model systems. Finally, in SA3, following autologous transplantation of aBMSCs into human craniofacial bone defects, we will determine how the phenotypic characteristics of different aBMSC populations correlate to their respective capacities to regenerate bone in vivo and clinically. Key outcome measures include: phenotypic characterization of aBMSCs; in vivo potential of aBMSCs to regenerate bone and vascular tissue; safety of aBMSC cell isolation and expansion methodologies for human application; clinical potential of aBMSCs to regenerate bone and vascular tissue; and examination of correlations between in vitro phenotype, in vivo bone regeneration, and clinical bone regeneration. Findings of the proposed study could significantly advance clinical cell therapy approaches for craniofacial regeneration. Additionally, the isolation of aBMSCs with the technique employed would have broad scientific impact in providing an alternative source of MSCs to be used in fundamental investigations of stem cell and bone biology.