ARTICLE | doi:10.20944/preprints202208.0122.v1
Subject: Medicine & Pharmacology, Other Keywords: neurogenic potential; neurites; SCAPs; sphere; stem cells
Online: 5 August 2022 (09:59:07 CEST)
Abstract: Cell-based neural regeneration is challenging due to the difficulty in obtaining sufficient neural stem cells with clinical applicability. SCAPs originating from embryonic neural crest with high neurogenic potential could be a promising cell source for neural regeneration. This study aimed to investigate whether the formation of 3D spheres can promote SCAPs neurogenic potential. Material and methods: 3D SCAPs spheres were first generated in 256-well agarose microtissue mold. The spheres and single cells were individually cultured on collagen I coated μ-Slide for 4 and 7 days. Cell morphological changes, neural marker expression, and neurite outgrowth were evaluated under a confocal microscope. Secretion of BDNF and NGF-β was measured by ELISA kits. Results: Pronounced morphological changes were noticed in a time-dependent manner. The migrating cells’ morphology changed from fibroblast-like cells to neuron-like cells. Compared to the 2D culture, neurite length, number, and the expression of neural markers, including Nestin, β-tubulin III, NeuN, and MAP-2 were significantly increased in the 3D spheres, while the secretion of BDNF and NGF-β was markedly downregulated at day 7. Conclusion: The formation of 3D spheres enhanced the neurogenic potential of SCAPs, suggesting the advantage of using the 3D spheres of SCAPs for the treatment of neural diseases.
ARTICLE | doi:10.20944/preprints201907.0102.v1
Subject: Life Sciences, Biochemistry Keywords: microglia exosomes; 3D culture; proteomic study; glioma; neurites outgrowth
Online: 8 July 2019 (04:26:49 CEST)
Using a combination of pan proteomic platform associated with systemic biology analyses, we demonstrate that neonatal microglial cells derived from cortex and spinal cord expressed different phenotypes upon the physiological or pathological conditions. They also highlight great variability in protein production on both cellular and exosome levels. Bioinformatics data indicate for the cortical microglia anti-inflammatory and neurogenesis/tumorigenesis characteristics, while for the spinal cord microglia involvement in the inflammatory response. We confirmed these results by performing functional testing including neurite outgrowth assays in DRGs cell line, and glioma proliferation analysis in 3D spheroid cultures. Results from these in vitro assays indicate that the microglia located at different CNS areas reveal differential biological functions. While both microglia sources enhanced growth of DRGs axons, only the spinal microglia significantly attenuated glioma proliferation. Overall these findings are pointing to the fact that the origin of neonatal microglia affects the physio-pathological function, which may address the prevalence of the glioma in the brain in comparison with the spinal cord in adult.
ARTICLE | doi:10.20944/preprints202201.0174.v1
Subject: Life Sciences, Other Keywords: alpha-synuclein; post-translational modifications; Parkinson’s disease; Multiple system atrophy; Lewy bodies; Lewy neurites; Glial cytoplasmic inclusions; phosphorylation; nitration; immunohistochemistry
Online: 12 January 2022 (14:31:13 CET)
Aggregated alpha-synuclein (-synuclein) is the main component of Lewy bodies (LBs), Lewy neurites (LNs), and glial cytoplasmic inclusions (GCIs), which are pathological hallmarks of idiopathic Parkinson’s disease (IPD) and multiple system atrophy (MSA), respectively. Initiating factors that culminate in forming LBs/LNs/GCIs remain elusive. Several species of -synuclein exist, including phosphorylated and nitrated forms. It is unclear which -synuclein post-translational modifications (PTMs) appear within aggregates throughout disease pathology. Herein we aimed to establish the predominant synuclein PTMs in post-mortem IPD and MSA pathology using immunohistochemistry. We examined the patterns of three -synuclein PTMs (pS87, pS129, nY39) simultaneously in pathology-affected regions of 15 PD, 5 MSA, 6 neurologically normal controls. All antibodies recognized LBs, LNs, and GCIs, albeit to a variable extent. pS129 -synuclein antibody was particularly immunopositive for LNs and synaptic dot-like structures followed by nY39 -synuclein antibody. GCIs, neuronal inclusions, and small threads were positive for nY39 -synuclein in MSA. Quantification of the LB scores revealed that pS129 -synuclein was the dominant and earliest -synuclein PTM followed by nY39 -synuclein, while lower amounts of pSer87 -synuclein appeared later in disease progression in PD. These results may have implications for novel biomarker and therapeutic developments.