NEW SOURCE OF MULTIPOTENT MESENCHYMAL CELLS FOR TISSUE ENGINEERING STUDIES: WASTE HUMAN OVARIAN FOLLICULAR FLUID

3D biomaterials have been widely used in reconstructive bone surgery as cell carriers providing mechanical support and as promoters of cell attachment and proliferation. In particular, gelatine cryogel scaffolds are promising new biomaterials owing to their biocompatibility and to substain the differentiation of mesenchymal stromal stem cells (MSCs). MSCs are multipotent stem cells that proliferate onto the surfaces with fibroblastic morphology and can differentiate into osteoblasts, chondrocytes and adipocytes. These cells can be isolated from several sources, including bone marrow and adipose tissue. Our previously studies showed the possibility to obtain MSCs also from the human ovarian follicular liquid (FL) that is usually wasted during in vitro fertilization. In this study, our aim was to show the possibility to use FL cells combined to a promising biomaterials for tissue engineering in regenerative medicine. We tested the ability of these FL cells to grow and differentiate in minimal culture conditions on gelatine cryogel, in comparison with MSCs derived from human bone marrow. Samples and controls were analyzed with confocal and scanning electron microscopes. Results showed that mesenchymal FL cells on gelatine cryogel scaffold were able to adapt to surface structure of biomaterial scaffold, growing on gelatin cryogel not only on the top surface but also in the layers below till 60 μm of deepness. Positive immunostaining for vimentin and CD44 (a typical MSC marker) of these cells on cryogel confirmed their mesenchymal stemness and demonstrated an excellent compatibility with the biomaterial surface. Bone marrow MSCs, used as control, showed a similar behavior. Preliminary results showed also the capability of FL cells on scaffold to be induced to osteogenic differentiation, producing bone extracellular matrix and expressing some specific proteins (i.e.osteopontin). In conclusion, MSCs derived from waste human ovarian follicular liquid showed promising affinity with 3D gelatine cryogel, opening new potential developments in biotech and medical applications.