A novel efficient method to isolate human adipose-derived stromal cells from periumbilical biopsies without enzymatic digestion

Background: Among all the possible sources of mesenchymal stem cells, adipose tissue has raised great interest and has become one of the most investigated sources. Several protocols are available for isolating adipose-derived stromal cells (ASCs) and, recently, the International Fat Applied Technology Society (IFATS) highlighted the importance of standardization and harmonization of these procedures. The enzymatic digestion is among the most frequently used and it could represent a problematic issue for transfer to clinical applications. Aim: Set up a novel manual non-enzymatic method combined with the use of stem cells medium (SCM) for isolating ASCs from human periumbilical biopsies with a high yield and rapidity. Materials and Methods: ASCs isolated without enzymatic step and using SCM were cultured in both SCM and a-MEM, and characterized for plastic adherence, growth capability, expression of both mesenchymal markers and multipotency genes, and differentiation features into mesenchymal lineage. Results: By following our method, cells can be easily maintained in a-MEM after their isolation in SCM, and they fully adhere to the requirements defined in IFATS and the International Society for Cellular Therapy (ISCT) guidelines. Moreover, we also report that isolated ASCs express neural markers such as β-tubulin III, Nestin and glial fibrillary acidic protein (GFAP). Conclusions: Our method allows the quickly and easily ASCs isolation from human periumbilical biopsies. The obtained cells can be cultured in a commercially available medium without altering their mesenchymal properties. Moreover, the lack of the enzymatic step is a great advantage for all those translational preclinical studies that require a great amount of cells, without affecting their biological features.