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- W1582218935 abstract "Bone marrow mesenchymal stem cells (MSCs) have been shown to differentiate into pulmonary epithelial cells in animal models. To explore whether human MSCs can undergo differentiation into airway epithelia, which can be exploited as a novel gene therapy strategy for cystic fibrosis (CF), human MSCs expressing green florescent protein (GFP) were mixed with primary human airway epithelial cells in a ratio of 1:5, 1:10, or 1:20, and seeded on a semi-permeable filter membrane coated with human placenta collagen type VI. The cells were cultured at an air-liquid interface and were able to maintain the apical surface dry after one to two weeks with transepithelial resistance greater than 700 Ohms, suggesting the formation of tight junctions. Immunofluorescent staining with an antibody against cytokeratin 18 (CK18) demonstrated that the stem cells co-cultured with human airway cells expressed this epithelium-specific marker, whereas prior to the culture, there is no detectable CK18 expression by this approach. Morphologically, the GFP-labeled stem cells converted their original thin-spindle fibroblast shape or flat polygonal cell shape to a typical epithelial cuboidal shape with an accessory structure like cilia by confocal microscopy, suggesting that the hMSCs in the co-culture system transdifferentiate into airway epithelial cells. To confirm the finding, we further did immunostaining with an antibody against occludin, an epithelial tight junction protein. Flow cytometry and fluorescent microscopy showed that ~10% of the initially loaded GFP-MSCs express this critical epithelial marker. By cell sorting, we isolated the co-cultured hMSCs, and RT-PCR was performed with primers specific to human CFTR. The results demonstrated that the air-liquid culture induces hMSCs to express the CFTR gene. Furthermore, co-culture of normal hMSCs with CF airway epithelial cells resulted in the wild-type CFTR expression. More importantly, the MSCs isolated from CF patients were able to be gene-corrected. In vitro differentiation assays proved that the CFTR gene-corrected CF MSCs still possess their naive multipotency, such as differentiation along osteogenic, adipogenic, and chondrogenic pathways. The gene-corrected CF MSCs were able to contribute to apical Cl- secretion by chloride efflux assay in response to cAMP agonists. Conditions to increase the efficacy of MSC differentiation are currently under investigation. Therefore, we conclude that bone marrow mesenchymal stem cells may have potential applications in treating human airway diseases, such as CF." @default.
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- W1582218935 date "2004-05-01" @default.
- W1582218935 modified "2023-09-27" @default.
- W1582218935 title "Potential of Bone Marrow Mesenchymal Stem Cells for Cystic Fibrosis Therapy" @default.
- W1582218935 doi "https://doi.org/10.1016/j.ymthe.2004.06.455" @default.
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