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Mesenchymal cells are important components of specified niches in the lung, and can mediate a wide range of processes including tissue regeneration and repair. Dysregulation of these processes can lead to improper remodeling of tissue as observed in several lung diseases. The mesenchymal cells responsible remain poorly described, partially due to the heterogenic nature of the mesenchymal compartment and the absence of appropriate markers. Here, we describe that CD105
CD90
mesenchymal cells can be divided into two populations based on their expression of CD13/aminopeptidase N (CD105
CD90
CD13
and CD105
CD90
CD13
). By prospective isolation using FACS, we show that both these populations give rise to clonogenic fibroblast-like cells, but with an increased clonogenic and proliferative capacity of CD105
CD90
CD13
cells. Transcriptomic and spatial analysis pinpoints an adventitial fibroblast subset as the origin of CD105
CD90
CD13
clonogenic mesenchymal cells in human lung.