Expansion and characterization of human adipose derived stem cells inside the 3D micro-niche Nichoid

Stem cells (SCs) have shown great promises in the treatment of various diseases, especially neurodegenerative ones. Human Adipose Derived Stem Cells (hADSCs) represent a promising source for cell therapy, especially as their isolation is less invasive compared to the isolation from other organs or tissue. In the body, SCs reside in a specialized microenvironment, characterized by a unique combination of biophysical and biochemical properties. Together, these features allow the maintenance of stemness. Recently, biomaterials have been used to create 3D micro-scaffolds, which mimic the biomechanical characteristics of SC niches. In this context, the micro-niche “Nichoid” has shown the ability to induce pluripotency in SCs. The aim of this study was to investigate the proliferation and stemness properties of hADSCs expanded in this engineered niche. Nichoids were fabricated by laser two-photon polymerization onto glass coverslips using a biocompatible photoresist. hADSCs were isolated from lipoaspiarate adipose tissue obtained by elective liposuction performed in voluntary subjects with full knowledge of the events. Cells expanded inside the Nichoid have been characterized by viability assay, Real Time PCR and immunofluorescence analysis. Furthermore, adipose and neural differentiation were investigated. hADSCs grown inside the Nichoid show a significantly higher cell viability than in standard conditions. Furthermore, the expression of pluripotency markers was upregulated inside the Nichoid, suggesting that the 3D micro-scaffold permits the potentiation of stemness features. Together, these results demonstrate that the Nichoid drives stem cell pluripotency maintenance without any chemical media. Our findings represent a great promise for the Nichoid’s possible application in the field of regenerative medicine, in particular in cell transplantation in preclinical animal model of Parkinson’s disease and Spinal Cord Injury.