Fluorescence imaging of in vivo miR-124a-induced neurogenesis of neuronal progenitor cells using neuron-specific reporters

Abstract

BackgroundFacilitation of the differentiation of the stem cells toward neuronal lineage is crucial for enhancing the differentiation efficacy of grafted stem cells for the possible treatment of neurodegenerative disorders. MicroRNA124a (miR-124a) has been considered as a neuronal lineage regulator, possessing the capability to activate neuronal differentiation. In this study, using a neuronal promoter-based reporter and live-cell fluorescence imaging, we visualized in vitro and in vivo the enhanced neuronal differentiation of neuronal progenitor cells with miR-124a overproduction.MethodsThe neuron specific alpha1 tubulin promoter-driven RFP reporter (pTa1-RFP) was used to trace the miR-124a-induced neuronal differentiation in live cell condition. MiR-124a or miR-scramble in 10 % glucose buffer was mixed with in vivo-jetPEITM and in vivo fluorescence images were obtained daily using Maestro spectral fluorescent imager.ResultsNeurite outgrowth was clearly seen in F11 cells after miR-124a transfection, and immunofluorescence staining showed increase of Tuj1 and NF at 48 hours. When pTa1-RFP-transfected F11 cells were implanted simultaneously with miR-124a into the nude mice, gradually increasing reporter signals and morphological changes indicated neuronal differentiation for 48 hours in live cells in vitro. The miR-124a-treated F11 cells showed higher reporter signals on in vivo fluorescence imaging than miR-scramble-treated cells, which were verified by ex vivo confirmation of Tuj1 and NF expression.ConclusionsThese results indicated that neuronal reporter-based neurogenesis imaging can be used for monitoring miR-124a acting as neuronal activator when miRNA was injected in in vivo PEI-coated form for miRNA-mediated regenerative therapy.

Defining the role of 17β‐estradiol in human endometrial stem cells differentiation into neuron‐like cells

“…The data showed no considerable expression of GFAP (the astrocytic marker) and Olig2 (the oligodendrocytic marker). These data were in agreement with the findings of other research (Bianchi et al, 2018; Jang et al, 2016; Li et al, 2018; Liu & Hu, 2018).…”

“…These intermediate filaments are essential for the regulation of axonal diameter and radial growth during development. They can also affect the nerve conduction velocity or axonal transport by providing structural support for axons (Jang et al, 2016).…”

“…EnSCs, endometrial stem cells neural markers and its expression is increased during the early stages of neural differentiation. It is highly expressed in a number of neuronal precursors that are mitotically active and also in immature and postmitotic or differentiated neurons (Gürsel et al, 2011;Jang et al, 2016). thereafter, is continuously expressed at a low level (Yim et al, 2007).…”

miR‐19b‐3p induces cell proliferation and reduces heterochromatin‐mediated senescence through PLZF in goat male germline stem cells

“…Embedded blocks were sectioned at 2 µm. The protocol of immunofluorescence staining was performed according to the lab facilities and to work done by (Jang et al, ; Mu et al, ).…”

BMSCs and miR‐124a ameliorated diabetic nephropathy via inhibiting notch signalling pathway

“…Studies have found that islet function can be improved by the endogenesis islet cell regeneration. BMSCs have been a key cellular source for replacement therapy of DN in regenerative medicine [4][5][6][7] and it could be induced to differentiate islet-like cells 8,9 . BMSCs could migrate into a damaged kidney and regulate immune response.…”