Injured microenvironment directly guides the differentiation of engrafted Flk-1+ mesenchymal stem cell in lung

Objective. Time window is a key factor in the treatment of lung injury by mesenehymal stem cells (MSC) transplantation. This study was aimed to analyze the engraftment and differentiation behavior of MSC transplanted at different time points after lung irradiation, and the possible mechanisms were discussed. Materials and Methods. The thorax of C57BL/6 mice was exposed to 1400 cGy, then 1(+)MSCs from enhanced green fluorescent protein C57BL/6 mice were systemically injected into C57BL/6 mice at 4 hours, 60 days, and 120 days post thoracic exposure, respectively. The engraftment and differentiation of Flk-1(+)NISC transplanted at different time points were evaluated. Lung tissue was collected and analyzed for fibrosis. Expression of transforming growth factor (TGF)-beta 1 in the lung was qualified by semi-quantitative real-time reverse transcription polymerase chain reaction. In vitro, Flk-1(+)MSCs were cultured in epithelium induction media, together with damaged primary lung cells, supernatants of radiation-injured lung cells, or TGF-beta 1 to find the possible factors that might effect Flk-1(+)MSC differentiation. Results. Cells injected immediately after injury were shown to differentiate into functional lung cells, such as epithelial and endothelial cells. Cells injected 2 months later were mostly located in the interstitial area and appeared as myofibrocyte. The in vivo lung microenvironments at different time points after injury were different from each other, especially TGF-beta 1 expression. We demonstrated that cytokines secreted by irradiated lung cells could inhibit differentiation of Flk-1(+)MSCs into epithelial cells in vitro. Conclusions. Flk-1(+)MSCs injected into the lung immediately after irradiation could differentiate into functional lung cells, while those injected at later stage after irradiation would be involved in fibrosis development. Thus our in vivo and in vitro studies demonstrated that differentiation of Flk-1(+)MSCs is controlled by the microenvironment. (c) 2007 ISEH - Society for Hematology and Stem Cells. Published by Elsevier Inc.