CELL CORE

耶鲁大学(医学院和学院)的许多研究人员长期以来对 肌肉骨骼疾病。这些调查员代表许多部门和学科。这 对骨骼的病理生理学的兴趣已经创建了许多合作项目 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 他们自己实验室内的技术通常会导致对现有技术的昂贵复制 技术。虽然有标准的技术可用，但实验室内部在质量和数量上存在差异 细胞的死亡是不可避免的。从无数的骨细胞中分离骨细胞的困难加剧了这种变异。 基因工程小鼠模型现已问世。为了满足这一制度需求，CELL的核心是 作为YCCMD初步应用的一部分而开发。它仍然是五个主要组成部分之一 中心。作为细胞核心新举措的一部分，分离脂肪细胞前体和脂肪细胞 差异化协议现在可用。此外，还可以获得小鼠间充质干细胞 通过细胞核心。细胞核心的目标是促进真正的初级实验的使用， 分离的小鼠和大鼠成骨细胞、体外产生的破骨细胞样细胞、成骨前体细胞和骨 骨髓来源的脂肪细胞。作为我们成像计划的一部分，Cell Core已经将其服务扩展到 包括小动物的高分辨率射线照片。 该提案的新的具体目标是提供以下服务： 1.分离培养原代小鼠和胎鼠颅骨成骨细胞。这包括大规模的 为生化分析和RNA和DMA提纯做准备。 2.通过以下方法从小鼠骨髓或脾细胞中分化出破骨细胞样细胞：a)体外共培养 用成骨细胞或用肿瘤素-M或甲状旁腺激素诱导；b)用mcsf培养 和RANKL。 3.小鼠骨髓来源成骨细胞/脂肪细胞前体细胞(骨髓)的培养和分化 基质细胞) 4.原代培养小鼠骨髓间充质干细胞。 5.支持一个细胞库，该细胞库提供对大量经冷冻处理的细胞系的访问。 6.提供小动物骨密度测量(DXA)和X光片。 7.培训研究人员进行成骨细胞、破骨细胞和祖细胞的体外分离和培养 细胞，这样经过适当培训的研究人员就可以在他们的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单元，以便费用和重复 所付出的努力被最小化了。小组核心还将向试点项目和可行性项目提供支助，作为 我们继续努力吸引新的研究人员参与肌肉骨骼研究。核心已经建立了一个

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