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Journal: International Journal of Molecular Sciences
Article Title: Polarized Anti-Inflammatory Mesenchymal Stem Cells Increase Hippocampal Neurogenesis and Improve Cognitive Function in Aged Mice
doi: 10.3390/ijms24054490
Figure Lengend Snippet: Polarized PACAP-treated MSC maintain mesenchymal phenotype. ( A ). Immunophenotyping of PACAP-treated MSC in flow cytometry presents positive expression of the mesenchymal markers CD106 (>50%), CD29 (>80%), CD44 (>70%), CD73 (>60%) and sca-1 (>40%) but negative expression of the hematopoietic markers CD45 (<8%), and CD11b (<5%). Graphs represent flow cytometry histograms for the expression of the different markers. The negative control histogram is presented with the blue filled histogram. ( B ). Both naïve (N = 4) and PACAP-treated MSC (N = 3) expressed detectable levels of VPAC2 receptor mRNA, as detected in real-time PCR. Since the activation of Toll-like receptor 3 (TLR3) is an established marker of the MSC anti-inflammatory phenotype (MSC2) , we administered pituitary adenylate cyclase-activating peptide (PACAP), a neuropeptide with anti-inflammatory properties that is known to upregulate TLR3 and vice versa with TLR4, at 20 nM for 4 days to establish the anti-inflammatory MCS phenotype (MSC2). PACAP treatment of MSC in vitro (pMSC, N = 5) did not increase significantly the expression of TLR3 ( C ) or TLR4 ( D ) but increased the TLR3/TLR4 gene expression ratio ( E ) compared with naïve MSC (N = 4), as detected in real-time PCR. All graphs present mean ± SE. * p < 0.05, Student t -test. ns = non-significant.
Article Snippet:
Techniques: Flow Cytometry, Expressing, Negative Control, Real-time Polymerase Chain Reaction, Activation Assay, Marker, In Vitro, Gene Expression