15 research outputs found
Targeting the proteome of cellular fractions: focus on secreted proteins
The high complexity of the total cellular proteome underscores the need for a more targeted investigation of particular subcellular fractions as a means to detect the changes at the level of low abundance proteins. However, this approach requires the application of an enrichment strategy. In this chapter, we present the protocols, which have been used for the analysis of secretome from cell lines, targeting the investigation of protein expression changes
2016 Jun 15
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.We acknowledge the financial support of the Portuguese Foundation for Science and Technology (FCT) for a Ciencia 2007 program and IF Development Grant (AJS), a pre-doctoral fellowship to FGT (SFRH/69637/2010) and a PhD fellowship to SA (SFRH/BD/81495/2011), a Canada Research Chair in Biomedical Engineering (LAB) and a SSE Postdoctoral Fellowship (KMP). Premios Santa Casa Neurociencias - Prize Melo e Castro for Spinal Cord Injury Research. PEst-C/SAU/LA0001/2013-2014. Co-funded by "COMPETE Programa Operacional Factores de Competitividade", QREN, the European Union (FEDER - Fundo Europeu de Desenvolvimento Regional) and by The National Mass Spectrometry Network (RNEM) under the contract REDE/1506/REM/2005.info:eu-repo/semantics/publishedVersio