Secretome of Mesenchymal Progenitors from the Umbilical Cord Acts as Modulator of Neural/Glial Proliferation and Differentiation

Abstract

It was recently shown that the conditioned media (CM) of Human Umbilical Cord Perivascular Cells (HUCPVCs), a mesenchymal progenitor population residing within the Wharton Jelly of the umbilical cord, was able to modulate in vitro the survival and viability of different neuronal and glial cells populations. In the present work, we aimed to assess if the secretome of HUCPVCs is able to 1) induce the differentiation of human telencephalon neural precursor cells (htNPCs) in vitro, and 2) modulate neural/glial proliferation, differentiation and survival in the dentate gyrus (DG) of adult rat hippocampus. For this purpose, two separate experimental setups were performed: 1) htNPCs were incubated with HUCPVCs-CM for 5 days after which neuronal differentiation was assessed and, 2) HUCPVCs, or their respective CM, were injected into the DG of young adult rats and their effects assessed 7 days later. Results revealed that the secretome of HUCPVCs was able to increase neuronal cell differentiation in vitro; indeed, higher densities of immature (DCX(+) cells) and mature neurons (MAP-2(+) cells) were observed when htNPCs were incubated with the HUCPVCs-CM. Additionally, when HUCPVCs and their CM were injected in the DG, results revealed that both cells or CM were able to increase the endogenous proliferation (BrdU(+) cells) 7 days after injection. It was also possible to observe an increased number of newborn neurons (DCX(+) cells), upon injection of HUCPVCs or their respective CM. Finally western blot analysis revealed that after CM or HUCPVCs transplantation, there was an increase of fibroblast growth factor-2 (FGF-2) and, to a lesser extent, of nerve growth factor (NGF) in the DG tissue. Concluding, our results have shown that the transplantation of HUCPVCs or the administration of their secretome were able to potentiate neuronal survival and differentiation in vitro and in vivo.

Modulation of the Mesenchymal Stem Cell Secretome Using Computer-Controlled Bioreactors: Impact on Neuronal Cell Proliferation, Survival and Differentiation

“…3A–C,J). This was also noted (with BrdU positive cells) in one of our previous works where we compared injection of MSCs isolated from the umbilical cord tissue or their secretome (CM) into the DG13. Similar effects were also observed regarding neuronal differentiation.…”

“…Neurobasal-A medium was then added at the same ratio as the PPRF-msc6. Cells were placed in a humidified incubator at 37 °C and 5% CO 2 , for 24 h. Afterwards, the conditioned medium (CM) was collected as previously described13.…”

“…hNPCs were fixed, washed, and blocked as previously described13, being followed by a 1 h incubation (at 37 °C) with primary antibodies: nestin (1:200, Millipore, Billerica, MA, USA) to detect neural progenitors, rabbit anti-doublecortin (DCX; 1:500, Abcam, Cambridge, MA, USA) to detect immature neurons, mouse anti-rat microtubule associated protein-2 (MAP-2; 1:500, Sigma) and neuronal nuclei (NeuN; 1:100, Millipore) to detect mature neurons. Secondary antibodies namely, Alexa Fluor 488 goat anti-rabbit immunoglobulin G (IgG, Life Technologies) and Alexa Fluor 594 goat anti-mouse immunoglobulin G (IgG, Life Technologies) were used for 1 h at 37 °C and then 10 min with 4-6-diamidino-2-phenylindole-dihydrochloride (DAPI; Life Technologies).…”

“…Sections were permeabilized and washed as previously described13. For Ki-67 staining antigen retrieval was performed with citrate buffer treatment (15–20 min on microwave).…”

Manufacturing of Human Extracellular Vesicle-Based Therapeutics for Clinical Use

“…Rats that were injected with the secretome of hUC mesenchymal progenitors in the hippocampal neurogenic niche displayed similar levels of neural and glial proliferation and differentiation as those injected with parental cells. Moreover, the numbers of doublecortin positive neuronal progenitors in vivo either after MSC or MSC-EV injection were equally increased [56]. Finally, in a rat model of Parkinson’s disease, the secretome of BM-MSCs increased neurogenesis and cell survival and impacted on brain structure and animal behavior [57].…”