![]() P19 cells, a pluripotent embryonal cell line derived from mouse teratocarcinoma, can differentiate into cells of all three germ layers upon stimulation with various agents. In the present study, we used P19 cells to investigate the role of neuronal exosomes in differentiation of pluripotent P19 cells into neurons in the absence of B27 media supplement. RA is an inducer of neurogenesis and thus may have primed the cells for neuronal differentiation. Both studies utilized medium containing B27 that contains retinol, a precursor of retinoic acid (RA). In a 2 nd study, cyclin D1 enriched N2A neuronal exosomes induced neuron differentiation of mouse embryonic stem cells. The hiPSC-derived neuronal exosomes induce cell proliferation and neurogenesis through activation of signaling cascades. Two studies demonstrated neurogenic effects of neuronal exosomes. Neurons are terminally differentiated and are highly specialized cells. In addition, exosomes contain certain proteins and lipids that reflect their endosomal origin and are hence used as markers for their characterization. The composition of exosomal cargo changes with physiology and pathology of the parent cells. Exosomes exert their effects on recipient cells through their unique bioactive cargo of proteins, lipids, small RNAs, and/or metabolites that often reveal their cell origin (donor/parent cell). Exosomes are small microvesicles with a diameter ranging from ~ 30 to 150 nm originating in endosomes of cells and are released by all cells in vivo and in vitro. Intercellular communication through exosomes facilitates cell differentiation from progenitor/stem cells. Our novel findings on exosomes-mediated differentiation of UD-P19 to P19 neurons provide tools to study pathways directing neuron development/differentiation and develop novel therapeutic strategies in neuroscience. P19N exosomes provide an alternative method to genetic modifications for cellular differentiation of neurons. UD-P19 exosomes were rich with ncRNAs required for maintenance of stemness. Small RNA-seq identified enrichment of P19N exosomes with pro-neurogenic non-coding RNAs (ncRNAs) such as miR-9, let-7, MALAT1 and depleted with ncRNAs involved in maintenance of stem cell characteristics. Incubation with UD-P19 exosomes for six days did not affect UD-P19. Continuous exposure of UD-P19 to P19N exosomes for six days induced formation of small-sized embryoid bodies that differentiated into MAP2-/GluN2B-positive neurons recapitulating RA-induction of neurogenesis. ![]() P19N internalized significantly higher number of Dil-P19N exosomes as compared to UD-P19 with accumulation in the perinuclear region. Both UD-P19 and P19N released exosomes with characteristic exosome morphology, size, and common protein markers. Here we report P19N exosome-mediated differentiation of UD-P19 to P19N. Retinoic acid induces differentiation of P19 cells (UD-P19) to P19 neurons (P19N) that behave like cortical neurons and express characteristic neuronal genes such as NMDA receptor subunits. Exosomes play a role in tissue/organ development and differentiation. ![]()
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