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（类型2），用于从人类志愿者和啮齿动物的骨髓中制备成体干细胞/祖细胞，定义细胞的质量，然后将其分发给多个研究人员。我们制备的成体干细胞/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MSCs。自上次竞争性更新以来，我们向280多名科学家运送了627批1075瓶冷冻间充质干细胞，其中157人由18个NIH研究所资助。我们收到了140多封支持信。目标是：(1)继续生产人类间充质干细胞（hMSCs）的标准化制剂，以分发给其他研究者。我们将用慢病毒转导一些hMSCs，用细胞质绿色荧光蛋白（GFP）、线粒体红色荧光蛋白（mitred）和荧光素酶表达在培养和体内追踪细胞的有用基因。(2)继续提供类似的小鼠间充质干细胞（mMSCs）制剂，以分发给其他研究者。(3)继续提供具有相似特征的大鼠间充质干细胞（rMSCs）制备，供其他研究者使用。我们将继续从其他大鼠品系和普遍表达GFP的转基因大鼠中制备rmsc，作为培养和体内实验的标记物。(4)在探索性研究（占分配资金的10%）中，我们将使用微阵列、qRT-PCR和elisa检测分泌因子，比较不同标准制备的hMSCs和细胞因子预先激活的hMSCs表达的基因。将对这些数据进行分析，以确定抗炎和免疫调节基因表达水平最高的制剂。研究结果将提供给骨髓间充质干细胞的接受者，科学家们将被鼓励比较那些数据表明在他们正在测试的系统中可能存在疗效差异的制剂。我们将提供一名生物信息学合作专家的协助，以评估

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.