An Evaluation of the Stemness, Paracrine, and Tumorigenic Characteristics of Highly Expanded, Minimally Passaged Adipose-Derived Stem Cells

• Published in: PloS one 2016-09, Vol.11 (9), p.e0162332
• Main Authors: El Atat, Oula, Antonios, Diane, Hilal, George, Hokayem, Nabil, Abou-Ghoch, Joelle, Hashim, Hussein, Serhal, Rim, Hebbo, Clara, Moussa, Mayssam, Alaaeddine, Nada
• Format: Article
• Language: English
• Subjects: Adipocytes; Adipose tissue; Adipose Tissue - cytology; Adipose Tissue - immunology; Adolescent; Adult; Aldehyde dehydrogenase; Aldehyde Dehydrogenase - metabolism; Analysis; Apoptosis; Biology and Life Sciences; Bone marrow; c-Myc protein; Cancer; Carcinogenesis; Carcinogens; Cell culture; Cell Differentiation; Cell Lineage; Cytometry; Dehydrogenases; Dermatology; Diabetes; Differentiation; Enzyme-linked immunosorbent assay; Fibroblasts; Flow cytometry; Gene expression; Health aspects; Humans; Immunophenotyping; Inflammation; Interleukin 6; Kinases; Laboratories; MDM2 protein; Medicine; Medicine and Health Sciences; Mesenchymal stem cells; Middle Aged; Mutation; Myc protein; p53 Protein; Paracrine signalling; Physical characteristics; Plastic surgery; Prostaglandin E2; Regenerative medicine; Research and Analysis Methods; Secretions; Stem cells; Stem Cells - cytology; Stem Cells - immunology; Telomerase; Telomerase - metabolism; Therapeutic applications; Tissue engineering; Tissue inhibitor of metalloproteinase 2; Tumor proteins; Tumor suppressor genes; Tumor Suppressor Protein p53 - metabolism; Vascular endothelial growth factor; Young Adult
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0162332

The use of adipose-derived stem cells (ADSC) in regenerative medicine is rising due to their plasticity, capacity of differentiation and paracrine and trophic effects. Despite the large number of cells obtained from adipose tissue, it is usually not enough for therapeutic purposes for many diseases or cosmetic procedures. Thus, there is the need for culturing and expanding cells in-vitro for several weeks remain. Our aim is to investigate if long- term proliferation with minimal passaging will affect the stemness, paracrine secretions and carcinogenesis markers of ADSC. The immunophenotypic properties and aldehyde dehydrogenase (ALDH) activity of the initial stromal vascular fraction (SVF) and serially passaged ADSC were observed by flow cytometry. In parallel, the telomerase activity and the relative expression of oncogenes and tumor suppressor genes were assessed by q-PCR. We also assessed the cytokine secretion profile of passaged ADSC by an ELISA. The expanded ADSC retain their morphological and phenotypical characteristics. These cells maintained in culture for up to 12 weeks until P4, possessed stable telomerase and ALDH activity, without having a TP53 mutation. Furthermore, the relative expression levels of TP53, RB, and MDM2 were not affected while the relative expression of c-Myc decreased significantly. Finally, the levels of the secretions of PGE2, STC1, and TIMP2 were not affected but the levels of IL-6, VEGF, and TIMP 1 significantly decreased at P2. Our results suggest that the expansion of passaged ADSC does not affect the differentiation capacity of stem cells and does not confer a cancerous state or capacity in vitro to the cells.