Osteoprogenitor Subpopulations within Adipose-derived Stroma

脂肪源性基质内的骨祖细胞亚群

• 批准号: 7408828
• 负责人: Deepak M Gupta
• 金额: $ 5.06万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-03-29 至 2010-03-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7408828
• 关键词: Adipose tissue; Adult; Alkaline Phosphatase; Allogenic; Apatites; Attenuated; Biological Assay; CD44 gene; Calvaria; Cell Fractionation; Cell Lineage; Cell Separation; Cell surface; Cells; Cellular biology; Clinical; Data; Defect; Degenerative Disorder; Diagnostic radiologic examination; Disadvantaged; Discipline; Disease; ENG gene; Extracellular Matrix; FVB Mouse; Fatty acid glycerol esters; Fellowship; Flow Cytometry; Gene Expression; Harvest; Healed; Healthcare; Hematopoietic stem cells; Hereditary Disease; Histology; Human; In Vitro; Individual; Investigation; Knowledge; Masks; Medical; Modeling; Mus; Names; National Research Service Awards; Natural regeneration; Operative Surgical Procedures; Population; Primary Cell Cultures; Proliferating; Prosthesis; Proteins; Proto-Oncogene Protein c-kit; Research Infrastructure; Skeletal system; Skeleton; Sorting - Cell Movement; Staining method; Stains; Stem cells; Stromal Cells; Surface Antigens; Therapeutic; Time; Tissues; Week; Work; adult stem cell; alizarin; base; cell type; design; healing; in vitro Assay; in vivo; male; mature animal; multipotent cell; novel; postnatal; progenitor; repaired; research study; scaffold; size; skeletal regeneration; socioeconomics; tomography; type IB bone morphogenetic protein receptor

DESCRIPTION (provided by applicant): Adipose-derived stromal cells (ASCs) have the capacity to undergo osteoblastic differentiation. These postnatal cells may be used to design cell-based regenerative therapies for skeletal repair in debilitating genetic disorders, degenerative diseases, and traumatic or post-surgical tissue deficits. While we have made substantial progress in studying the cell biology underlying osteodifferentiation of ASCs, we are limited by working with a heterogeneous cell population, and specific enriched, subpopulations of osteoprogenitor cells with the greatest osteopotential need to be identified. Current investigations utilize heterogeneous cells and have several disadvantages. Subpopulation gene expression changes may be attenuated, or even completely masked, due to the presence of multiple cell types. Determining whether there is a single set of progenitor cells capable of osteodifferentiation would be extremely beneficial, both in terms of advancing our knowledge of postnatal multipotent cells and for optimizing clinical therapeutic strategies. Preliminary data suggest that it is indeed possible to enrich heterogeneous ASCs for osteoprogenitors using fluorescent-activated cell sorting (FACS). The first Specific Aim focuses on performing cell fractionation experiments on primary cultures of cells derived from inguinal fat pads of adult mice. Cells will be subdivided according to expression of individual candidate markers (i.e. marker+ vs. marker-) previously identified as markers of adult stem cells. These cells will be assayed in vitro for their potential to proliferate and differentiate down the osteogenic lineage compared to unfractionated ASCs. In the second Specific Aim, the ASC subpopulations identified in the first Specific Aim with greatest osteopotential will be placed in an in vivo mouse calvarial defect model to determine if they enhance skeletal regeneration. Together, these Aims will use flow-cytometry in concert with both in vitro and in vivo osteoblastic differentiation assays to prospectively delineate the cell-surface marker profile of osteoprogenitors residing within the heterogeneous mix of harvested ASCs. Skeletal disease poses a serious challenge to a variety of medical disciplines. At the same time, it also poses a significant socioeconomic burden on our healthcare infrastructure exceeding $1 billion annually. Current clinical approaches to regeneration of the skeleton include use of autogenous grafts, allogenic materials and prosthetic products, each with its own inherent disadvantages. The ability to harness the regenerative potential of cellular building block progenitor cells portends an exciting and novel paradigm for management of skeletal disease.

描述（由申请人提供）：脂肪源性基质细胞（ASC）具有进行成骨细胞分化的能力。这些出生后细胞可用于设计基于细胞的再生疗法，用于修复衰弱性遗传性疾病、退行性疾病以及创伤或手术后组织缺陷的骨骼。虽然我们在研究 ASC 骨分化的细胞生物学方面取得了实质性进展，但我们受到异质细胞群研究的限制，并且需要鉴定具有最大骨潜能的特定富集骨祖细胞亚群。目前的研究利用异质细胞并存在一些缺点。由于多种细胞类型的存在，亚群基因表达变化可能会减弱，甚至完全被掩盖。确定是否存在一组能够骨分化的祖细胞将非常有益，无论是在增进我们对出生后多能细胞的了解方面，还是在优化临床治疗策略方面。初步数据表明，使用荧光激活细胞分选（FACS）确实可以富集骨祖细胞的异质 ASC。第一个具体目标侧重于对源自成年小鼠腹股沟脂肪垫的细胞的原代培养物进行细胞分级分离实验。将根据先前确定为成体干细胞标记的各个候选标记（即标记+与标记-）的表达来细分细胞。与普通 ASC 相比，这些细胞将在体外进行增殖和分化成骨谱系的潜力进行分析。在第二个特定目标中，第一个特定目标中鉴定出的具有最大骨潜能的 ASC 亚群将被置于体内小鼠颅骨缺损模型中，以确定它们是否增强骨骼再生。总之，这些目标将使用流式细胞术与体外和体内成骨细胞分化测定相结合，前瞻性地描绘存在于收获的 ASC 的异质混合物中的骨祖细胞的细胞表面标记谱。骨骼疾病对多种医学学科提出了严峻的挑战。与此同时，它还给我们的医疗基础设施带来每年超过 10 亿美元的巨大社会经济负担。目前骨骼再生的临床方法包括使用自体移植物、同种异体材料和假体产品，每种方法都有其固有的缺点。利用细胞构建块祖细胞的再生潜力的能力预示着骨骼疾病管理的令人兴奋的新颖范例。