CELL CORE

细胞核心

• 批准号: 8376753
• 负责人: MARK C HOROWITZ
• 金额: $ 18.98万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-04-01 至 2014-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8376753
• 关键词: Address; Adipocytes; Animals; Biochemical; Biology; Bone Marrow; Bone Marrow Cells; CSF1 gene; Calvaria; Cell Count; Cell Culture Techniques; Cell Line; Cells; Coculture Techniques; Cost Savings; Densitometry; Dual-Energy X-Ray Absorptiometry; Engineering; Freezing; Functional disorder; Genotype; Goals; Hormones; Image; In Vitro; Laboratories; Macrophage Colony-Stimulating Factor; Marrow; Mesenchymal Stem Cells; Modeling; Mus; Musculoskeletal; Musculoskeletal Diseases; Osteoblasts; Osteoclasts; Parathyroid gland; Phenotype; Preparation; Protocols documentation; RNA; Rattus; Research; Research Personnel; Resolution; Services; Skeleton; Spleen; Stem cells; Stromal Cells; TNFSF11 gene; Techniques; Technology; Training; Universities; Variant; adipocyte differentiation; base; bone; bone cell; cell bank; college; fetal; interest; investigator training; medical schools; member; oncostatin M; progenitor; skeletal

Many investigators at Yale University (Medical School and College) have a long-standing interest in musculoskeletal disorders. These investigators represent numerous departments and discjplines. This interest in the pathophysiology of the skeleton has created a number of collaborative projects among interested investigators. As a result of these interactions, investigators have sought to develop cell culture techniques within their own laboratories often leading to expensive duplication of already existing technologies. Although standard techniques are available, intra-laboratpry variation in the quality and number of cells is unavoidable. This variation is exacerbated by the difficulties in isolating bone cells from the myriad of genetically engineered murine models now available. To address this Institutional need, the Cell Core was developed as part of the initial application of the YCCMD. It remains one of the five major components of the Center. As part of the Cell Core's new initiatives, isolation of adipocyte precursors and adipocyte differentiation protocols are now available. In addition, murine mesenchymal stem cells may be obtained through the Cell Core. The goal of the Cell Core is to facilitate the experimental use of authentic primary, isolated mouse and rat osteoblasts, osteoclast-like cells generated in vitro, osteoblast precursors, and bone marrow-derived adipocytes. As part of our imaging initiative, the Cell Core has expanded its services to include small animal high-resolution radiographs. The new Specific Aims of the proposal are to provide the following services: 1. Isolate and culture primary murine and fetal rat calvarial osteoblasts. This includes large-scale preparations for biochemical analyses and RNA and DMA purification. 2. Differentiate murine osteoclast-like cells from bone marrow or spleen cells by: a) in vitro coculture with osteoblasts or induction with Oncostatin-M or parathyroid hormone; b) culture with MCSF and RANKL. 3. Grow and differentiate mouse bone marrow derived osteoblast/adipocyte progenitors (marrow stromal cells) 4. Supply primary mouse mesenchymal stem cells (MSCs). 5. Support a cell bank that provides access to a large number of cryoperserved cell lines. 6. Provide small animal bone densitometry (DXA) and radiographs. 7. Train investigators for in vitro isolation and culture of osteoblasts, osteoclasts and progenitor cells, so that investigators appropriately trained can establish these techniques in their laboratories The Core will provide cells to Center members in a coordinated manner, such that expenses and duplication of effort are minimized. The Cell Core will also provide support to the Pilot and Feasibility Projects as part of our continuing effort to attract new investigators to musculoskeletal research. The Core has an established

耶鲁大学（医学院和学院）的许多研究人员长期以来一直对以下方面感兴趣： 肌肉骨骼疾病这些调查人员代表了许多部门和学科。这 对骨骼的病理生理学的兴趣已经创造了许多合作项目， 感兴趣的调查员。作为这些相互作用的结果，研究人员试图开发细胞培养物， 在他们自己的实验室里使用这些技术，往往会导致昂贵的重复， 技术.尽管有标准的技术，但实验室内的质量和数量 细胞是不可避免的。这种变异由于难以从大量的骨细胞中分离出骨细胞而加剧。 基因工程小鼠模型的研究。为了满足这一机构需求， 作为YCCMD初始应用的一部分开发。它仍然是五个主要组成部分之一， 中心作为Cell Core新举措的一部分，分离脂肪细胞前体和脂肪细胞 分化方案现在是可用的。此外，可以获得鼠间充质干细胞， 穿过细胞核心细胞核心的目标是促进实验使用真实的主要， 分离的小鼠和大鼠成骨细胞、体外产生的破骨细胞样细胞、成骨细胞前体和骨 骨髓来源的脂肪细胞。作为我们成像计划的一部分，Cell Core已将其服务扩展到 包括小动物高分辨率射线照片。 该提案新的具体目标是提供以下服务： 1.原代分离培养小鼠和胎鼠颅骨成骨细胞。其中包括大规模 用于生化分析和RNA和DMA纯化的制备。 2.通过以下方法从骨髓或脾细胞中分化小鼠破骨细胞样细胞：a）体外共培养 用成骨细胞或用抑瘤素-M或甲状旁腺激素诱导; B）用MCSF培养 和RANKL。 3.小鼠骨髓源性成骨/脂肪祖细胞（骨髓 基质细胞） 4.提供原代小鼠间充质干细胞（MSCs）。 5.支持一个细胞库，提供大量冷冻保存细胞系。 6.提供小动物骨密度测定（DXA）和X光片。 7.培养研究者进行成骨细胞、破骨细胞和祖细胞的体外分离和培养 细胞，以便经过适当培训的研究人员可以在实验室中建立这些技术 核心将以协调的方式向中心成员提供细胞， 的努力被最小化。该小组核心还将为试点和可行性项目提供支持， 我们不断努力吸引新的研究人员到肌肉骨骼研究。核心有一个既定的