Use of preparations comprising exosomes derived from mesenchymal stem cells (MSCs) in the prevention and therapy of inflammatory conditions
The present invention relates to the use of exosome-preparations derived from neonatal or adult tissue-derived mesenchymal stem-cells (MSCs) for the prevention or for therapy of inflammatory conditions, such as, for example, pre- and postnatally acquired damages of the brain (i.e. neuronal damages) or in complications following stem cell transplantation (“graft vs host-disease”, GvHD).
1. A method for producing a pharmaceutical preparation comprising exosomes that exhibit an immunomodulatory effect, wherein said immunomodulatory effect is selected from a reduced IL-1β, a reduced TNF-α and a reduced IFN-γ cytokine response of effectors peripheral blood mononuclear cells (PBMC) of a recipient of said preparation, said method comprising the following steps:
a) providing a cell culture medium supernatant from neonatal or adult tissue-derived mesenchymal stem-cells (MSCs) that have been cultured in a medium supplemented platelet lysate, the cell culture supernatant comprising exosomes,
b) enriching said exosomes via a method selected from at least one of differential centrifugation, filtration, polyethylene glycol precipitation and/or monolithic chromatography,
c) quantifying said exosomes and analyzing at least one of the specific markers selected from CD63, CD81 and tumor susceptibility gene 101 (Tsg101), in said enriched exosomes,
(d) determining an in vitro anti-inflammatory effect and/or an immune suppressive effect of said enriched exosomes by detecting at least one marker selected from anti-inflammatory molecules IL-10, TGF-β1 and HLA-G in said enriched exosomes,
(e) determining a reduced IL-1β, a reduced TNF-α and/or a reduced IFN-γ cytokine response of donor effector PBMC cells incubated with said enriched exosomes,
f) selecting those enriched exosomes that exhibit said anti-inflammatory effect and/or immune suppressive effect, wherein the levels of TGF-β1 as measured in an activity test are at least ten times higher than in plasma levels of healthy controls and wherein the ratio of IL-10 to IFN-γ is more than 1.
2. The method according to claim 1 , wherein said preparation as produced is suitable for intravenous administration.