Differentiation of Adult Stem Cells to Lung Ex Vivo

成体干细胞离体向肺的分化

基本信息

批准号：
7463755
负责人：
DARWIN Johnson PROCKOP
金额：
$ 36.69万
依托单位：
TULANE UNIVERSITY OF LOUISIANA
依托单位国家：
美国
项目类别：
财政年份：
资助国家：
美国
起止时间：
至
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/7463755
关键词：
Address Alveolar Animal Model Animals Biological Assay Cell fusion Cells Class Clinical Medicine Clinical Trials Coculture Techniques Disease model Endothelium Engraftment Epithelial Cells Epithelium Genes Green Fluorescent Proteins Heat-Shock Response Human Hyperoxia Ice Immunodeficient Mouse Immunohistochemistry Infusion procedures Laboratories Lentivirus Vector Lung Marrow Mesenchymal Stem Cells Molecular Mononuclear Mus Personal Satisfaction Polymerase Chain Reaction Population Preparation Rattus Reporting Research Personnel Sampling Series Single Nucleotide Polymorphism Standards of Weights and Measures Stromal Cells Subfamily lentivirinae System Testing Time Tissues Vascular Endothelial Cell Western Blotting Wound Healing adult stem cell blastomere structure cell injury experience human disease immunocytochemistry injured monolayer promoter repaired research study self renewing cell

项目摘要

We will use a recently developed co-culture system and an assay for competitive engraftment to identify the best candidatesfor use. The Specific Aims are: (1.) Use standard and unfractionated preparations of human and rat MSCs in the recently developed co-culture system with small airway epithelial cells (SAECs) damaged by heat shock or hyperoxia to generate quantitative assays for (a) differentiation of MSCs; (b) fusion of MSCs with SAECs; and (c) the fate of MSCs that fuse with SAECs. (2.) Use the co-culture assay developed in Specific Aim 1 to determine which of three sub-populations of MSCs that have recently been characterized in our laboratory are the most effective in differentiating into SAECs: (a) small and rapidly self-renewing cells (RS cells); (b) large and slowly replicating mature MSCs (mMSCs); and (c) cells that we have recently isolated and tentatively referred to as pre-RS cells because they are more similar to embryonic cells than RS cells. To define molecular mechanisms that explain any observed differences among cells in the assay, we will carry out microarray assays on the cells before and after addition to the co-cultures. (3.) Use the co-culture assay from Specific Aim 1 to test the same populations of MSCs in analogous co-culture systems in which the target cells are large bronchial epithelial cells, alveolar epithelial cells, or pulmonary vascular endothelial cells. (4.) Determine which of the subpopulations of MSCs from Specific Aim 2 can most efficiently engraft in lung by using a competitive engraftment assay in immunodeficient mice.

我们将使用最近开发的共培养系统和竞争性植入的测定法，以识别最佳候选人使用。具体目的是：（1。）在最近开发的共培养系统中使用人类和大鼠MSC的标准和未分流的制剂与热休克或高氧损坏的小气道上皮细胞（SAEC）来产生（a）MSC分化的定量测定；（b）MSC与SAEC的融合；（c）与SAEC融合的MSC的命运。（2.）使用特定目标1中开发的共培养测定法确定最近在我们的实验室中表征的三个亚群中的哪个子群是最有效的区分SAEC：（a）小而快速的自我更新细胞（RS细胞）；（b）大而缓慢复制成熟的MSC（MMSC）；（c）我们最近分离并暂定地称为前RS细胞的细胞，因为它们与胚胎细胞更相似，而不是RS细胞。为了定义分子机制来解释分析中细胞中观察到的任何差异，我们将在添加共培养之前和之后对细胞进行微阵列测定。（3.）使用特定目标1的共培养测定法测试类似的共培养系统中相同的MSC群体，其中靶细胞是大支气管上皮细胞，肺泡上皮细胞或肺血管内皮细胞。（4.）通过在免疫缺陷型小鼠中使用竞争性植入测定法，确定特定目标2中MSC的哪些亚群可以最有效地植入肺中。