title: Unbiased and quantitative proteomics reveals highly increased angiogenesis induction by the secretome of mesenchymal stromal cells isolated from fetal rather than adult skin.
abstract: Scarless wound healing and functional regeneration are typical processes of the fetus, gradually lost during postnatal life, and maximally attributed to fetal skin tissue and induced by fetal skin fibroblasts. The latter have been successfully applied to postnatal wounds, with clear advantages compared with autologous dermis grafts or adult fibroblast applications. Our goal was to functionally identify and uncover key factors and mechanisms through the analysis of secretomes, the principal players in all cell therapies based on mesenchymal stromal cells (MSCs). Cell secretomes also putatively mediate skin regenerative effects achieved in clinical applications of fetal skin fibroblasts. An innovative and unbiased approach of comparative and quantitative proteomics of cell conditioned media enabled us to gain knowledge of key molecules and processes from a translational perspective. Using banks of fetal and adult skin fibroblasts that we previously characterized as being MSCs, we discovered secretome changes by identification and comparative quantification, distinguishing secretome signatures of fetal skin MSCs putatively relevant for therapeutic microenvironment modulation. The uncovered proteins can trigger, directly and by modulation of extracellular matrix, angiogenesis, thus highlighting its key role towards scarless wound healing. The angiogenic trigger was functionally validated and corroborated in vitro, with fetal skin MSC secretomes stabilizing and inducing the formation of capillary-like networks by endothelial cells and fetal liver MSCs, respectively. Our approach and our results may aid in the development of cell-based and cell-free products for skin regeneration in acute or chronic injury, and also for wound healing in the regeneration of other tissues. Copyright © 2017 John Wiley & Sons, Ltd.
authors.affiliation: Fondazione Ri.MED, Palermo, Italy.|Regenerative Medicine and Biomedical Technologies Unit, Department of Laboratory Medicine and Advanced Biotechnologies, IRCCS-ISMETT (Mediterranean Institute for Transplantation and Advanced Specialized Therapies), Palermo, Italy.
authors.forename: Massimiliano
authors.identifier: 0000-0001-5610-0797
authors.initials: M
authors.lastname: Gaetani
authors.affiliation: Fondazione Ri.MED, Palermo, Italy.|Regenerative Medicine and Biomedical Technologies Unit, Department of Laboratory Medicine and Advanced Biotechnologies, IRCCS-ISMETT (Mediterranean Institute for Transplantation and Advanced Specialized Therapies), Palermo, Italy.
authors.forename: Cinzia Maria
authors.initials: CM
authors.lastname: Chinnici
authors.affiliation: Fondazione Ri.MED, Palermo, Italy.|Regenerative Medicine and Biomedical Technologies Unit, Department of Laboratory Medicine and Advanced Biotechnologies, IRCCS-ISMETT (Mediterranean Institute for Transplantation and Advanced Specialized Therapies), Palermo, Italy.
authors.forename: Anna Paola
authors.initials: AP
authors.lastname: Carreca
authors.affiliation: Fondazione Ri.MED, Palermo, Italy.|Regenerative Medicine and Biomedical Technologies Unit, Department of Laboratory Medicine and Advanced Biotechnologies, IRCCS-ISMETT (Mediterranean Institute for Transplantation and Advanced Specialized Therapies), Palermo, Italy.
authors.forename: Claudia
authors.initials: C
authors.lastname: Di Pasquale
authors.affiliation: Fondazione Ri.MED, Palermo, Italy.|Regenerative Medicine and Biomedical Technologies Unit, Department of Laboratory Medicine and Advanced Biotechnologies, IRCCS-ISMETT (Mediterranean Institute for Transplantation and Advanced Specialized Therapies), Palermo, Italy.
authors.forename: Giandomenico
authors.initials: G
authors.lastname: Amico
authors.affiliation: Fondazione Ri.MED, Palermo, Italy.|Regenerative Medicine and Biomedical Technologies Unit, Department of Laboratory Medicine and Advanced Biotechnologies, IRCCS-ISMETT (Mediterranean Institute for Transplantation and Advanced Specialized Therapies), Palermo, Italy.
authors.forename: Pier Giulio
authors.initials: PG
authors.lastname: Conaldi
mesh_terms: D000328:Adult; D053687:Adult Stem Cells / Q000166:cytology* / Q000378:metabolism; D002469:Cell Separation; D020405:Dermis / Q000166:cytology; D053686:Fetal Stem Cells / Q000166:cytology* / Q000378:metabolism; D005786:Gene Expression Regulation; D006801:Humans; D016130:Immunophenotyping; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000378:metabolism; D039902:Multipotent Stem Cells / Q000166:cytology / Q000378:metabolism; D018919:Neovascularization, Physiologic; D020543:Proteome / Q000378:metabolism*; D040901:Proteomics / Q000379:methods*; D012867:Skin / Q000166:cytology*; D014945:Wound Healing
chemical_list: D020543:Proteome
keywords: angiogenesis; fetal skin MSCs; liquid chromatography tandem mass spectrometry; microenvironment modulation; quantitative proteomics; secretome; stable isotope labelling by amino acids in cell culture; tissue regeneration