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

Abstract

In recent years it has been shown that the therapeutic benefits of human mesenchymal stem/stromal cells (hMSCs) in the Central Nervous System (CNS) are mainly attributed to their secretome. The implementation of computer-controlled suspension bioreactors has shown to be a viable route for the expansion of these cells to large numbers. As hMSCs actively respond to their culture environment, there is the hypothesis that one can modulate its secretome through their use. Herein, we present data indicating that the use of computer-controlled suspension bioreactors enhanced the neuroregulatory profile of hMSCs secretome. Indeed, higher levels of in vitro neuronal differentiation and NOTCH1 expression in human neural progenitor cells (hNPCs) were observed when these cells were incubated with the secretome of dynamically cultured hMSCs. A similar trend was also observed in the hippocampal dentate gyrus (DG) of rat brains where, upon injection, an enhanced neuronal and astrocytic survival and differentiation, was observed. Proteomic analysis also revealed that the dynamic culturing of hMSCs increased the secretion of several neuroregulatory molecules and miRNAs present in hMSCs secretome. In summary, the appropriate use of dynamic culture conditions can represent an important asset for the development of future neuro-regenerative strategies involving the use of hMSCs secretome.

Secretome of Undifferentiated Neural Progenitor Cells Induces Histological and Motor Improvements in a Rat Model of Parkinson's Disease

“…Finally, Dickkopf 3, was another molecule identified in the secretome of hNPCs, which has been described as an important modulator of DAergic neuronal differentiation through the Wnt/β‐catenin signaling pathway . Still, as previously stated by Teixeira and colleagues , all the mentioned proteins have already been identified in different cell secretomes by distinct proteomic‐based techniques, supporting the results stated by our proteomic analysis. Hereupon, our findings suggest that the modulation effect in DA neurons triggered by hNPCs secretome could be related with the presence/expression of several secreted factors, as those discussed above.…”

Impact of the Secretome of Human Mesenchymal Stem Cells on Brain Structure and Animal Behavior in a Rat Model of Parkinson's Disease

“…Additionally, a specific library of precursor masses and fragment ions created from a pool of hMSC samples, described by Teixiera et al , was used to performed protein identification from the SWATH analysis. The chromatographic profiles of the peptides presented in the MSC library were extracted from the SWATH‐mass spectrometry data using the SWATH processing plug‐in for PeakView (version 2.0.01; SCIEX).…”

Ultrafiltration Segregates Tissue Regenerative Stimuli Harboured Within and Independent of Extracellular Vesicles

“…Indeed, the microenvironment of culture 61 and/or tissue source of origin [62][63][64][65] can lead to dynamic phenotypes and functional characteristics demonstrated by MSC in vitro 58,66 . It is also becoming evident that serial passaging and unwanted differentiation of MSC can lead to a loss of therapeutic activity 14,19,67 , such as the acquisition of a senescent secretome 68 .…”

“…A collection of studies has demonstrated the potential of utilizing MSC as EV-generating biofactories and engineering the secretome of MSC to meet therapeutic needs 37,41,67 . Our previous studies have supported this progression of thought and we envision the creation of a cell-free biotherapeutics 'tailored' to combat a given pathology.…”