Unveiling the effects of the secretome of mesenchymal progenitors from the umbilical cord in different neuronal cell populations

Abstract

It has been previously shown that the secretome of Human Umbilical Cord Perivascular Cells (HUCPVCs), known for their mesenchymal like stem cell character, is able to increase the metabolic viability and hippocampal neuronal cell densities. However, due to the different micro-environments of the distinct brain regions it is important to study if neurons isolated from different regions have similar, or opposite, reactions when in the presence HUCPVCs secretome (in the form of conditioned media-CM). In this work we: 1) studied how cortical and cerebellar neuronal primary cultures behaved when incubated with HUCPVCs CM and 2) characterized the differences between CM collected at two different conditioning time points. Primary cultures of cerebellar and cortical neurons were incubated with HUCPVCs CM (obtained 24 and 96 hours after three days of culturing). HUCPVCs CM had a higher impact on the metabolic viability and proliferation of cortical cultures, than the cerebellar ones. Regarding neuronal cell densities it was observed that with 24h CM condition there were higher number MAP-2 positive cells, a marker for fully differentiated neurons; this was, once again, more evident in cortical cultures. In an attempt to characterize the differences between the two conditioning time points a proteomics approach was followed, based on 2D Gel analysis followed by the identification of selected spots by tandem mass spectrometry.Results revealed important differences in proteins that have been previously related with phenomena such as neuronal cell viability, proliferation and differentiation, namely 14-3-3, UCHL1, hsp70 and peroxiredoxin-6. In summary, we demonstrated differences on how neurons isolated from different brain regions react to HUCPVCs secretome and we have identified different proteins (14-3-3 and hsp70) in HUCPVCs CM that may explain the above referred results M

The Secretome of Bone Marrow and Wharton Jelly Derived Mesenchymal Stem Cells Induces Differentiation and Neurite Outgrowth in SH-SY5Y Cells

“…As it can be observed, BMSCs CM 24 h as well as HUCPVCs CM 24 h and 96 h had very similar results to those of the RA-differentiated cells group ( P > 0.05) (Figure 4 and Figures 5(a) to 5(c) and 5(e)), which is a strong indicator of the differentiation effects of the secretome of both BMSCs and HUCPVCs. Finally, the decrease in the mean neurite length observed in the BMSCs CM 96 h group ( P < 0.05) (Figure 4 and Figures 5(a) and 5(d)) may be related with the half-life of neurotrophic factors present in the CM at the point of their collection [19, 36, 37]. …”

“…Indeed, SH-SY5Y cells differentiated with HUCPVCs CM 96 h exhibited significant greater expression of both DRD2 and DAT genes than cells differentiated with BMSCs CM 96 h ( P < 0.05, P < 0.01). The later result suggests not only that the different secretome composition of different tissue derived MSCs induces SH-SY5Y cells differentiation into different neuronal phenotypes but also that the effects mediated by the secretome of MSCs in neuronal differentiation are associated with the temporal profile of CM collection [19, 36, 37]. …”

“…CM was collected from P6 BMSCs and HUCPVCs as previously reported by Fraga et al [37]. Shortly, cells were plated at a density of 4.000 cells/cm 2 and allowed to grow for 3 days in a 5% humidified CO 2 atmosphere at 37°C.…”

“…For instance, Ribeiro et al [36] and Fraga et al [37] revealed that the secretome of mesenchymal progenitors isolated from the Wharton's jelly of the umbilical cord increased neuronal cell viability and cell densities. These effects were attributed to the expression of NGF and the vesicular fraction of the secretome, respectively, which contained proteins typically involved in neuroprotection.…”

Hydrogels and Cell Based Therapies in Spinal Cord Injury Regeneration

“…In another study, both adipose stem cells (ASCs) and human umbilical cord perivascular cells (HUCPVCs) CMs were shown to increase hippocampal neurons survival and metabolic activity [73]. More recently, the secretome of HUCPVCs was also found to increase cell viability, proliferation, and neuronal cell densities in both cortical and cerebellar neuronal cultures [74]. Other in vitro and in vivo studies showed similar results [75–77].…”