Presently, we implicate progranulin and decorin as additional paracrine factors that exert an anti-apoptotic effect against A42-neurotoxicity (Figure ?(Figure4).4). for rat primary neuronal cells did not react with decorin and Nesbuvir progranulin of hUCB-MSCs. These data suggest that secretion of decorin and progranulin were induced in hUCB-MSCs by the co-culture of rat primary neuronal cells in the presence or absence of A42. Open in a separate window Figure 1 Decorin and progranulin are highly secreted from human umbilical cord blood-derived mesenchymal stem cells. A: Human umbilical cord blood-derived mesenchymal stem cells (hUCB-MSCs) were co-cultured with amyloid-42-exposed rat primary neuronal cells for 24 h in a Transwell chamber. Then, rat primary neuronal cells were stained by Live/Dead staining. Green color indicates surviving neuronal cells. B: Percentage of dead cells was calculated ( 0.05, = 4 per group); C: To identify paracrine factors, co-cultured media was analyzed by antibody-based array (RayBio). Spot intensity of progranulin and decorin in co-cultured media was much higher compared to rat primary neuronal cells in the absence of hUCB-MSC ( 0.05, = 3 per group); D: Each medium used in the Transwell was analyzed by enzyme-linked immunosorbant assay for progranulin and decorin (a 0.05, c 0.05, = 3 per group). Treatment of recombinant decorin and progranuiln increases neuron viabilty To test whether decorin and progranulin participate Rabbit Polyclonal to MSK2 in the neuroprotection against A42-neurotoxicity, recombinant Nesbuvir decorin and progranulin were treated in A42-exposed rat primary neurons at three doses (10 ng/mL, 20 ng/mL and 50 ng/mL). After treatment of recombinant decorin or progranulin for 36 h, rat primary neuronal cells were analyzed by Live/Dead staining. Almost the same neuroprotective effect of decorin and progranulin was apparent at each dose (data not shown). Representative percentage of dead cells using 10 ng of decorin or progranulin in A42-exposed rat primary neuronal cells is shown in Figure ?Figure2B.2B. Since neuron and glia cells were mixed in the rat primary neuronal cells, we tried to stain MAP2-positive neurons in recombinant decorin- and progranuin-treated cells exposed to A42 (Figure ?(Figure2C).2C). Treatment of each protein reduced A-mediated neurotoxicity because MAP2-positive cells were very apparent in A-exposed neurons with decorin or progranulin, compared to controls. These data suggest that secreted decorin and progranulin from hUCB-MSCs have an anti-apoptotic effect against A42-neurotoxicity 0.05, = 3 per group) and decorin (B) (a 0.05, = 3 per group); C: Rat primary neurons in each condition were stained by anti-microtubule associated protein 2 (MAP2) antibody. Red color indicates MAP2-positive neurons and nuclei were visualized by DAPI (blue); D: The percentage of MAP2-postive neuron in each conditions were analyzed (a 0.05, = 3 per group). CONT: Control; PGRN: Progranulin; DCN: Decorin. DISCUSSION MSC display paracrine action in pathological conditions. Especially, we observed that hUCB-MSCs also secreted a variety of proteins by incubation with body fluid collected from patients (unpublished data). When we analyzed co-cultured media by various biochemical approaches such as antibody array, we identified several proteins including galectin-3 and sICAM-1. We have previously reported on the function of each identified protein in Nesbuvir an AD model[14,15]. Here, we report on the role of decorin and progranulin in hUCB-MSC-action in an AD model. hUCB-MSCs seem to act simultaneously in an AD microenviroment because A reduction and increased neuron survival were observed after application of the hUCB-MSCs. During co-culture of hUCB-MSCs with rat primary neurons, secretion of decorin and progranulin were increased in the presence or absence Nesbuvir of A42 (Figure ?(Figure1).1). These elevations were confirmed by two-different methods: antibody-based array and ELISA. Since hUCB-MSCs protected from A42 neurotoxicity a direct effect on neurons. Decorin pretreatment of meningial fibroblasts resulted in a three-fold increase in neurite outgrowth from co-cultured adult sensory.