TY - JOUR
T1 - Protein and Molecular Characterization of a Clinically Compliant Amniotic Fluid Stem Cell-Derived Extracellular Vesicle Fraction Capable of Accelerating Muscle Regeneration Through Enhancement of Angiogenesis
AU - Mellows, Ben
AU - Mitchell, Robert
AU - Antonioli, Manuela
AU - Kretz, Oliver
AU - Chambers, David
AU - Zeuner, Marie Theres
AU - Denecke, Bernd
AU - Musante, Luca
AU - Ramachandra, Durrgah L.
AU - Debacq-Chainiaux, Florence
AU - Holthofer, Harry
AU - Joch, Barbara
AU - Ray, Steve
AU - Widera, Darius
AU - David, Anna L.
AU - Huber, Tobias B.
AU - Dengjel, Joern
AU - De Coppi, Paolo
AU - Patel, Ketan
N1 - Funding Information:
This research was funded by BBSRC, National Institute for Health Research Great Ormond Street Biomedical Research Centre, and Rosetrees Trust. A.L.D. is funded by the National Institute for Health Research University College London Hospitals Biomedical Research Centre. P.D.C. is funded by the National Institute for Health Research and Great Ormond Street Hospital Children’s Charity. T.B.H. was supported by the DFG (CRC1140, CRC 992, HU 1016/ 8-1), by the BMBF (01GM1518C), by the European Research Council-ERC grant 616891, and by the H2020-IMI2 consortium BEAt-DKD. D.W. was supported by a grant from the DFG (WI4318/2-1).
Publisher Copyright:
© Copyright 2017, Mary Ann Liebert, Inc. 2017.
Copyright:
Copyright 2018 Elsevier B.V., All rights reserved.
PY - 2017/9/15
Y1 - 2017/9/15
N2 - The secretome of human amniotic fluid stem cells (AFSCs) has great potential as a therapeutic agent in regenerative medicine. However, it must be produced in a clinically compliant manner before it can be used in humans. In this study, we developed a means of producing a biologically active secretome from AFSCs that is free of all exogenous molecules. We demonstrate that the full secretome is capable of promoting stem cell proliferation, migration, and protection of cells against senescence. Furthermore, it has significant anti-inflammatory properties. Most importantly, we show that it promotes tissue regeneration in a model of muscle damage. We then demonstrate that the secretome contains extracellular vesicles (EVs) that harbor much, but not all, of the biological activity of the whole secretome. Proteomic characterization of the EV and free secretome fraction shows the presence of numerous molecules specific to each fraction that could be key regulators of tissue regeneration. Intriguingly, we show that the EVs only contain miRNA and not mRNA. This suggests that tissue regeneration in the host is mediated by the action of EVs modifying existing, rather than imposing new, signaling pathways. The EVs harbor significant anti-inflammatory activity as well as promote angiogenesis, the latter may be the mechanistic explanation for their ability to promote muscle regeneration after cardiotoxin injury.
AB - The secretome of human amniotic fluid stem cells (AFSCs) has great potential as a therapeutic agent in regenerative medicine. However, it must be produced in a clinically compliant manner before it can be used in humans. In this study, we developed a means of producing a biologically active secretome from AFSCs that is free of all exogenous molecules. We demonstrate that the full secretome is capable of promoting stem cell proliferation, migration, and protection of cells against senescence. Furthermore, it has significant anti-inflammatory properties. Most importantly, we show that it promotes tissue regeneration in a model of muscle damage. We then demonstrate that the secretome contains extracellular vesicles (EVs) that harbor much, but not all, of the biological activity of the whole secretome. Proteomic characterization of the EV and free secretome fraction shows the presence of numerous molecules specific to each fraction that could be key regulators of tissue regeneration. Intriguingly, we show that the EVs only contain miRNA and not mRNA. This suggests that tissue regeneration in the host is mediated by the action of EVs modifying existing, rather than imposing new, signaling pathways. The EVs harbor significant anti-inflammatory activity as well as promote angiogenesis, the latter may be the mechanistic explanation for their ability to promote muscle regeneration after cardiotoxin injury.
KW - miRNA
KW - muscle
KW - regeneration
KW - secretome
UR - http://www.scopus.com/inward/record.url?scp=85029223185&partnerID=8YFLogxK
U2 - 10.1089/scd.2017.0089
DO - 10.1089/scd.2017.0089
M3 - Article
AN - SCOPUS:85029223185
SN - 1547-3287
VL - 26
SP - 1316
EP - 1333
JO - Stem Cells and Development
JF - Stem Cells and Development
IS - 18
ER -