Human Adipose Tissue-Derived SSEA-4 Subpopulation Multi-Differentiation Potential Towards the Endothelial and Osteogenic Lineages

• Type: Article in International Scientific Journal
• Date: 2013

Authors

Mihaila, SM (Author) Other; Frias, AM (Author) Other; Pirraco, RP (Author) Other; Rada, T (Author) Other; Rui Reis (Author) Other; Gomes, ME (Author) Other; Marques, AP (Author) Other

Journal

Title: Tissue Engineering - Part A

Vol. 19

Pages: 235-246

ISSN: 1937-3341

Publisher: Mary Ann Liebert

Indexing

ISI Web of Knowledge - 40 Citations

Scopus - 41 Citations

Other information

Authenticus ID: P-002-23S

DOI: 10.1089/ten.tea.2012.0092

Abstract (EN): 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 alpha-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 low-density lipoprotein and to form capillary-like structures when seeded on Matrigel. SSEA-4(+)hASCs cultured in alpha-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.

Language: English

Type (Professor's evaluation): Scientific

No. of pages: 12