Differentiation of Adult Stem Cells to Lung Ex Vivo

我们将使用最近开发的共培养系统和竞争移植试验来确定最佳候选者。具体目标是：(1)；在最近开发的与热休克或高氧损伤的小气道上皮细胞（SAECs）共培养系统中，使用标准和未分离的人和大鼠间充质干细胞制剂进行定量分析(a)间充质干细胞的分化；(b) MSCs与saec的融合；(c)与saec融合的MSCs的命运。(2)。使用在Specific Aim 1中开发的共培养试验来确定最近在我们实验室中表征的MSCs的三个亚群中哪一个是最有效的

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.