Modulation properties of factors released by bone marrow stromal cells on activated microglia: an in vitro study

Abstract

In the present paper we develop a new non-cell based (cell-free) therapeutic approach applied to BV2 microglial cells and spinal cord derived primary microglia (PM) using conditioned media from rat bone marrow stromal cells (BMSCs-CM). First we collected conditioned media (CM) from either naive or injured rat spinal cord tissue (SCI-CM, inflammatory stimulation agent) and from rat bone marrow stromal cells (BMSCs-CM, therapeutic immunomodulation agent). They were both subsequently checked for the presence of chemokines and growth, neurotrophic and neural migration factors using proteomics analysis. The data clearly showed that rat BMSCs-CM contain in vitro growth factors, neural migration factors, osteogenic factors, differentiating factors and immunomodulators, whereas SCI-CM contain chemokines, chemoattractant factors and neurotrophic factors. Afterwards we determined whether the BMSCs-CM affect chemotactic activity, NO production, morphological and pro-apoptotic changes of either BV2 or PM cells once activated with SCI-CM. Our results confirm the anti-migratory and NO-inhibitory effects of BMSCs-CM on SCI-CM-activated microglia with higher impact on primary microglia. The cytotoxic effect of BMSCs-CM occurred only on SCI-CM-stimulated BV2 cells and PM, not on naive BV2 cells, nor on PM. Taken together, the molecular cocktail found in BMSCs-CM is favorable for immunomodulatory properties.

Polysaccharides from Ganoderma lucidum attenuate microglia-mediated neuroinflammation and modulate microglial phagocytosis and behavioural response

“…In addition, we also recorded during LPS activation BV2 cells elongated and extended from the mostly short and compact morphology to a long shape. This finding coincides with a previous report in which BV2 cells were incubated in the conditioned medium from spinal cord injury for 3–48 h [49]. We provided quantitative descriptions for changes in morphology as decreased cell circularity with increased Feret’s diameter and confirmed that the changes ultimately led to the augmented cell area and perimeter.…”

Exploiting the impact of the secretome of MSCs isolated from different tissue sources on neuronal differentiation and axonal growth

“…Some other factors not related to MSC so far but that were recently shown to have regenerative and neurotrophic functions are the ASC-secreted Macrophage-colony stimulating factor (MCSF), Matrix metalloproteases (MMPS), Follistatin (FST)-like 1, Mesencephalic astrocyte-derived neurotrophic factor (MANF), and Neuron derived neurotrophic factor (NDNF) [38]. Others like Semaphorins (SEM), Galectins (Gal), Platelet-derived growth factor (PDGF) and Transforming growth factor-beta (TGF-b) were found in the secretome of BMSCs by Cizkova et al [39]. Altogether, these results suggest the existence of tissue-source based differences, as recently evidenced by Pires et al [28].…”

Coenzyme Q10 Regulation of Apoptosis and Oxidative Stress in H2O2 Induced BMSC Death by Modulating the Nrf-2/NQO-1 Signaling Pathway and Its Application in a Model of Spinal Cord Injury

“…BMSCs at a density of 3 × 10 5 cells were used for the phenotypic analysis using flow cytometry [26]. In brief, the cells were stained with phycoerythrin-(PE-) labeled primary antibodies against CD11, CD73, and the corresponding isotype control antibody of rat IgG1, in the dark at room temperature, and then fixed in 1% paraformaldehyde (PFA), after being washed twice with ice-cold PBS.…”