Preparation and Distribution of Adult Stem Cells

描述（由申请人提供）：这是一份竞争性更新申请（2型），申请中心从人类志愿者和啮齿动物的骨髓中制备成体干/祖细胞，确定细胞的质量，然后将其分发给多名研究者。我们制备的成体干/祖细胞最初被称为成纤维细胞集落形成单位，然后在血液学文献中被称为骨髓基质细胞，随后被称为间充质干细胞，最近被称为多能间充质基质细胞或MSC。自上次竞争性更新以来，我们向280多名科学家运送了627批1，075瓶冷冻MSC，其中157人由18个NIH研究所资助。我们收到了140多封支持信。目的：（1）继续生产标准化的人骨髓间充质干细胞（hMSCs）制剂，供其他研究者使用。我们将用慢病毒感染一些hMSCs，以表达在培养和体内追踪细胞中有用的基因：细胞质绿色荧光蛋白（GFP），线粒体红色荧光蛋白（MitoRed）和荧光素酶。(2)继续向其他研究者提供类似的小鼠间充质干细胞（mMSCs）制剂。(3)继续提供具有相似特征的大鼠间充质干细胞（rMSCs）制剂，以分发给其他研究者。我们将继续从其他大鼠品系和从普遍表达GFP的转基因大鼠中制备rMSCs，作为培养和体内实验的标记。(4)在探索性研究中（高达10%的拨款），我们将使用微阵列，qRT-PCR和分泌因子的ELISA a比较不同标准hMSCs制剂和细胞因子预活化培养的hMSCs表达的基因。将分析数据以鉴定具有最高水平的抗炎和免疫调节基因表达的制剂。研究结果将提供给hMSC的接受者，并鼓励科学家比较数据表明在他们测试的系统中可能存在疗效差异的制剂。我们将提供生物信息学合作专家的协助， 数据

DESCRIPTION (provided by applicant): This a competing renewal application (Type 2) for a Center to prepare adult stem/progenitor cells from the bone marrow of human volunteers and rodents, define the quality of the cells, and then distribute them to multiple investigators. The adult stem/progenitor cells we prepare were originally referred to as fibroblastic colony-forming-units, then as marrow stromal cells in the hematological literature, subsequently as mesenchymal stem cells, and most recently as multipotent mesenchymal stromal cells or MSCs. Since the last competitive renewal, we made 627 shipments of 1,075 vials of frozen MSCs to over 280 scientists of whom 157 are funded by 18 NIH Institutes. We have received over 140 Letters of Support. The Aims are: (1) To continue to produce standardized preparations of human MSCs (hMSCs) for distribution to other investigators. We will transduce some of the hMSCs with lentiviruses to express genes useful in tracking the cells both in culture and in vivo: with cytoplasmic green fluorescent protein (GFP), mitochondrial red fluorescent protein (MitoRed) and luciferase. (2) To continue to provide similar preparations of mouse MSCs (mMSCs) for distribution to other investigators. (3) To continue to provide similarly characterized preparations of rat MSCs (rMSCs) for distribution to other investigators. We will continue to prepare rMSCs from other rat strains and from transgenic rats ubiquitously expressing GFP as a marker for experiments in culture and in vivo. (4) In exploratory research (up to 10% of allocated funds), we will compare the genes expressed by different standard preparations of hMSCs and hMSCs pre-activated in culture by cytokines using microarrays, qRT-PCR and ELISAa for secreted factors. The data will be analyzed to identify the preparations with the highest levels of expression of anti-inflammatory and immune regulatory genes. The results will be provided to recipients of the hMSCs and scientists will be encouraged to compare preparations that the data suggest may vary in efficacy in the systems they are testing. We will provide assistance from a collaborating expert in bioinformatics in evaluating the data.