Evaluation of the multipotent character of human adipose tissue-derived stem cells isolated by Ficoll gradient centrifugation and red blood cell lysis treatment

Abstract

In the present study, the multipotent potential of two differential isolated human adipose-derived stem cell (hADSC) populations was evaluated. More specifically, hADSC isolated by means of classical Ficoll (F) gradient centrifugation were compared to hADSC isolated by means of red blood cell (RBC) lysis treatment and subsequent cultivation as 3D spheres. No significant difference in the genotypic expression of the multipotent markers Oct-4, Sox-2, Nanog, Klf-4 and cMyc could be observed between both isolation methods. Upon adipogenic and osteogenic differentiation, both hADSC populations showed lipid droplet accumulation and mineral deposition, respectively. Although, a more pronounced mineral deposition was observed in hADSC-RBC, suggesting a higher osteogenic potential. Upon exposure to keratinogenic media, both hADSC populations expressed the keratinocyte markers filaggrin and involucrin, evidencing a successful keratinogenic differentiation. Yet, no differences in expression were observed between the distinctive isolation procedures. Finally, upon exposure to neurogenic differentiation media, a significant difference in marker expression was observed. Indeed, hADSC-RBC only expressed vimentin and nestin, whereas hADSC-F expressed vimentin, nestin, NF-200, MBP and TH, suggesting a higher neurogenic potential. In summary, our data suggest that the choice of the most efficient isolation procedure of hADSC depends on the differentiated cell type ultimately required.