title: Angiogenic and anti-inflammatory properties of micro-fragmented fat tissue and its derived mesenchymal stromal cells.
abstract: BACKGROUND
  Adipose-derived mesenchymal stromal cells (Ad-MSCs) are a promising tool for advanced cell-based therapies. They are routinely obtained enzymatically from fat lipoaspirate (LP) as SVF, and may undergo prolonged ex vivo expansion, with significant senescence and decline in multipotency. Besides, these techniques have complex regulatory issues, thus incurring in the compelling requirements of GMP guidelines. Hence, availability of a minimally manipulated, autologous adipose tissue would have remarkable biomedical and clinical relevance. For this reason, a new device, named Lipogems® (LG), has been developed. This ready-to-use adipose tissue cell derivate has been shown to have in vivo efficacy upon transplantation for ischemic and inflammatory diseases. To broaden our knowledge, we here investigated the angiogenic and anti-inflammatory properties of LG and its derived MSC (LG-MSCs) population.
  
  
  METHODS
  Human LG samples and their LG-MSCs were analyzed by immunohistochemistry for pericyte, endothelial and mesenchymal stromal cell marker expression. Angiogenesis was investigated testing the conditioned media (CM) of LG (LG-CM) and LG-MSCs (LG-MSCs-CM) on cultured endothelial cells (HUVECs), evaluating proliferation, cord formation, and the expression of the adhesion molecules (AM) VCAM-1 and ICAM-1. The macrophage cell line U937 was used to evaluate the anti-inflammatory properties, such as migration, adhesion on HUVECs, and release of RANTES and MCP-1.
  
  
  RESULTS
  Our results indicate that LG contained a very high number of mesenchymal cells expressing NG2 and CD146 (both pericyte markers) together with an abundant microvascular endothelial cell (mEC) population. Substantially, both LG-CM and LG-MSC-CM increased cord formation, inhibited endothelial ICAM-1 and VCAM-1 expression following TNFα stimulation, and slightly improved HUVEC proliferation. The addition of LG-CM and LG-MSC-CM strongly inhibited U937 migration upon stimulation with the chemokine MCP-1, reduced their adhesion on HUVECs and significantly suppressed the release of RANTES and MCP-1.
  
  
  CONCLUSIONS
  Our data indicate that LG micro-fragmented adipose tissue retains either per se, or in its embedded MSCs content, the capacity to induce vascular stabilization and to inhibit several macrophage functions involved in inflammation.
authors.affiliation: Cellular Neurobiology Laboratory, Department of Cerebrovascular Diseases, IRCCS Neurological Institute C. Besta, Via Celoria 11, 20131 Milan, Italy.
authors.forename: Valentina
authors.initials: V
authors.lastname: Ceserani
authors.affiliation: Cellular Neurobiology Laboratory, Department of Cerebrovascular Diseases, IRCCS Neurological Institute C. Besta, Via Celoria 11, 20131 Milan, Italy.
authors.forename: Anna
authors.initials: A
authors.lastname: Ferri
authors.affiliation: Department of Clinical and Experimental Sciences, Institute of Pathological Anatomy, University of Brescia, Brescia, Italy.
authors.forename: Angiola
authors.initials: A
authors.lastname: Berenzi
authors.affiliation: Department of Clinical and Experimental Sciences, Institute of Pathological Anatomy, University of Brescia, Brescia, Italy.
authors.forename: Anna
authors.initials: A
authors.lastname: Benetti
authors.affiliation: Laboratory of Clinical Pathology and Neurogenetic Medicine, Fondazione IRCCS Neurological Institute C. Besta, Milan, Italy.
authors.forename: Emilio
authors.initials: E
authors.lastname: Ciusani
authors.affiliation: Department of Veterinary Medicine, University of Perugia, Perugia, Italy.
authors.forename: Luisa
authors.initials: L
authors.lastname: Pascucci
authors.affiliation: Department of Veterinary Medicine, University of Perugia, Perugia, Italy.
authors.forename: Cinzia
authors.initials: C
authors.lastname: Bazzucchi
authors.affiliation: Department of Biomedical, Surgical and Dental Sciences, University of Milan, Milan, Italy.
authors.forename: Valentina
authors.initials: V
authors.lastname: Coccè
authors.affiliation: Department of Biomedical, Surgical and Dental Sciences, University of Milan, Milan, Italy.
authors.forename: Arianna
authors.initials: A
authors.lastname: Bonomi
authors.affiliation: Department of Biomedical, Surgical and Dental Sciences, University of Milan, Milan, Italy.
authors.forename: Augusto
authors.initials: A
authors.lastname: Pessina
authors.affiliation: San Michele Veterinary Hospital, Tavezzano con Villavesco, Lodi, Italy.
authors.forename: Erica
authors.initials: E
authors.lastname: Ghezzi
authors.affiliation: San Michele Veterinary Hospital, Tavezzano con Villavesco, Lodi, Italy.
authors.forename: Offer
authors.initials: O
authors.lastname: Zeira
authors.affiliation: Department of Veterinary Medicine, University of Perugia, Perugia, Italy.
authors.forename: Piero
authors.initials: P
authors.lastname: Ceccarelli
authors.affiliation: Image Institute, Milan, Italy.
authors.forename: Silvia
authors.initials: S
authors.lastname: Versari
authors.affiliation: Image Institute, Milan, Italy.
authors.forename: Carlo
authors.initials: C
authors.lastname: Tremolada
authors.affiliation: Cellular Neurobiology Laboratory, Department of Cerebrovascular Diseases, IRCCS Neurological Institute C. Besta, Via Celoria 11, 20131 Milan, Italy.
authors.forename: Giulio
authors.initials: G
authors.lastname: Alessandri