CELL CORE

细胞核心

基本信息

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
项目类别：
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

项目摘要

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）的许多调查人员对肌肉骨骼疾病。这些调查人员代表了许多部门和脱节。这对骨骼的病理生理学的兴趣已创造了许多协作项目有兴趣的调查员。由于这些相互作用，研究人员试图发展细胞培养他们自己的实验室中的技术通常会导致已经存在的昂贵重复技术。尽管可以使用标准技术，但质量和数字的内部差异细胞不可避免。从无数的骨细胞隔离骨细胞的困难使这种变化加剧了现在可以使用基因工程的鼠模型。为了满足这一机构需求，细胞核心是开发为YCCMD初始应用的一部分。它仍然是中心。作为细胞核心新计划的一部分，脂肪细胞前体和脂肪细胞的隔离现在可以使用差异化协议。此外，可以获得鼠间充质干细胞通过细胞核。细胞核心的目的是促进真实主要的实验使用，分离的小鼠和大鼠成骨细胞，骨细胞状细胞在体外产生，成骨细胞前体和骨骼骨髓来源的脂肪细胞。作为我们成像计划的一部分，细胞核心将其服务扩展到包括小动物高分辨率X光片。该提案的新特定目的是提供以下服务： 1。孤立和培养的原代鼠和胎儿大鼠颅骨成骨细胞。这包括大规模生化分析以及RNA和DMA纯化的准备。 2。通过以下方式将鼠骨细胞样细胞与骨髓或脾细胞区分开：a）体外共培养与骨细胞或抗科斯汀-M或甲状旁腺激素诱导； b）与MCSF的文化和Rankl。 3。生长和区分小鼠骨髓衍生成骨细胞/脂肪细胞祖细胞（骨髓）基质细胞） 4。供应原代小鼠间充质干细胞（MSC）。 5。支持一个细胞库，该单元格提供访问大量冷冻的细胞系。 6。提供小动物骨密度测定法（DXA）和X光片。 7。训练研究人员，用于体外隔离和成骨细胞，骨细胞和祖细胞的培养细胞，以便经过适当培训的研究人员可以在其实验室中建立这些技术核心将以协调的方式为中心成员提供细胞，以便支出和重复努力被最小化。电池核心还将为飞行员和可行性项目提供支持我们持续的努力吸引新的研究人员进行肌肉骨骼研究。核心已建立