Stromal Vascular Fraction Cell Sheets Angiogenic Potential for Tissue Engineering and Regenerative Medicine Applications

last updated: 2015-09-08
TitleStromal Vascular Fraction Cell Sheets Angiogenic Potential for Tissue Engineering and Regenerative Medicine Applications
Publication TypeConference Abstract -ISI Web of Science Indexed
Year of Publication2015
AuthorsCosta M., Cerqueira M. T., Rodrigues D. B., Santos T. C., Marques A. P., Pirraco R. P., and Reis R. L.

One of the biggest concerns in the Tissue Engineering field is the correct vascularization of engineered constructs. Strategies involving the use of endothelial cells are promising but adequate cell sourcing and neo-vessels stability are enduring challenges. In this work, we propose the hypoxic pre-conditioning of the stromal vascular fraction (SVF) of human adipose tissue to obtain highly angiogenic cell sheets (CS). For that, SVF was isolated after enzymatic dissociation of adipose tissue and cultured until CS formation in normoxic (pO2=21%) and hypoxic (pO2=5%) conditions for 5 and 8 days, in basal medium. Immunocytochemistry against CD31 and CD146 revealed the presence of highly branched capillary-like structures, which were far more complex for hypoxia. ELISA quantification showed increased VEGF and TIMP-1 secretion in hypoxia for 8 days of culture. In a Matrigel assay, the formation of capillary-like structures by endothelial cells was more prominent when cultured in conditioned medium recovered from the cultures in hypoxia. The same conditioned medium increased the migration of adipose stromal cells in a scratch assay, when compared with the medium from normoxia. Histological analysis after implantation of 8 days normoxic- and hypoxic-conditioned SVF CS in a hindlimb ischemia murine model showed improved formation of neo-blood vessels. Furthermore, Laser Doppler results demonstrated that the blood perfusion of the injured limb after 30 days was enhanced for the hypoxic CS group.

Overall, these results suggest that SVF CS created under hypoxia can be used as functional vascularization units for tissue engineering and regenerative medicine.

JournalTissue Engineering Part A
Conference Name2015 4th TERMIS World Congress
Date Published2015-08-28
PublisherMary Ann Liebert, Inc. publishers
Conference LocationBoston, Massachusetts
KeywordsIn vivo, regenerative medicine, Tissue engineering
Peer reviewedno

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