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TitleHuman adipose tissue-derived SSEA-4 subpopulation multi-differentiation potential towards the endothelial and osteogenic lineages
Author(s)Mihaila, Silvia M.
Frias, A. M.
Pirraco, Rogério
Rada, Tommaso
Reis, R. L.
Gomes, Manuela E.
Marques, A. P.
KeywordsEndothelial and osteogenic differentiation
SSEA-4+ human adipose derived stem/stromal cells
Issue date2013
PublisherMary Ann Liebert
JournalTissue Engineering : part A
Abstract(s)Human adipose tissue has been recently recognized as a potential source of stem cells for regenerative medicine applications, including bone tissue engineering (TE). Despite the gathered knowledge regarding the differentiation potential of human adipose tissue-derived stem cells (hASCs), in what concerns the endothelial lineage many uncertainties are still present. The existence of a cell subpopulation within the human adipose tissue that expresses a SSEA-4 marker, usually associated to pluripotency, raises expectations on the differentiation capacity of these cells (SSEA-4+hASCs). In the present study, the endothelial and osteogenic differentiation potential of the SSEA-4+hASCs was analyzed, aiming at proposing a single-cell source/ subpopulation for the development of vascularized bone TE constructs. SSEA-4+hASCs were isolated using immunomagnetic sorting and cultured either in a-MEM, in EGM-2 MV (endothelial growth medium), or in osteogenic medium. SSEA-4 +hASCs cultured in EGM-2 MV formed endothelial cell-like colonies characterized by a cobblestone morphology and expression of CD31, CD34, CD105, and von Willebrand factor as determined by quantitative reverse transcriptase (RT)-polymerase chain reaction, immunofluorescence, and flow cytometry. The endothelial phenotype was also confirmed by their ability to incorporate acetylated lowdensity lipoprotein and to form capillary-like structures when seeded on Matrigel. SSEA-4 +hASCs cultured in a-MEM displayed fibroblastic-like morphology and exhibited a mesenchymal surface marker profile (>90% CD90+/CD73+/CD105+). After culture in osteogenic conditions, an overexpression of osteogenic-related markers (osteopontin and osteocalcin) was observed both at molecular and protein levels. Matrix mineralization detected by Alizarin Red staining confirmed SSEA-4 +hASCs osteogenic differentiation. Herein, we demonstrate that from a single-cell source, human adipose tissue, and by selecting the appropriate subpopulation it is possible to obtain microvascular-like endothelial cells and osteoblasts, the most relevant cell types for the creation of vascularized bone tissue-engineered constructs.
Publisher version
AccessRestricted access (UMinho)
Appears in Collections:3B’s - Artigos em revistas/Papers in scientific journals

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