Nicotinamide adenine dinucleotide rejuvenates septic bone marrow mesenchymal stem cells

Figure 1 Morphology, clonogenic capacity, proliferation and characterization of bone marrow mesenchymal stem cells. A: Representative images depicting the morphology of in vitro-expanded bone marrow mesenchymal stem cells (BMSCs) from healthy donors (HD) and patients with sepsis of various generations; B: Representative images of colony-forming units-fibroblast assays, with results expressed as the number of colony-forming units-fibroblast per one million bone marrow mononuclear cells for HD samples (n = 14) and septic samples (n = 15); C-E: Proliferation of HD and septic BMSCs, C: Merged channel image showing EdU (magenta) and DAPI (blue) staining and the percentage of EdU-positive cells relative to the total number of cells in HD BMSCs (n = 10) and septic BMSCs (n = 10) (C). Growth curves were assessed using the CCK-8 assay for HD BMSCs (n = 10) and septic BMSCs (n = 10) (D). Expression levels of proliferating cell nuclear antigen detected by quantitative real-time polymerase chain reaction (HD: n = 13; sepsis: n = 10) (E); F: Representative flow cytometry images illustrating typical BMSC markers. Positive markers: CD73, CD90, CD105; negative markers: CD34 and CD45. Each error bar represents the mean ± SEM. P values were determined using a t-test (^aP < 0.05, ^cP < 0.001). HD: Healthy donors; CFU-F: Colony-forming units-fibroblast; PCNA: Proliferating cell nuclear antigen.