Interaction of Human, Canine and Murine Adipose-Derived Stem Cells with Different Biomaterials

摘要

Due to ease of collection and in vitro expansion, abundance and plasticity, adipose-derived stem cells (ADSC) represent an attractive option for cell therapy and tissue engineering. A variety of biodegradable scaffolds are used as substitutes for the artificial extracellular matrix. Among them are biomaterials consisting of biomolecules such as collagen and poly (L-lactic acid) (OPLA), and bioceramics such as calcium phosphate. As an advantage compared to conventional two-dimensional (2D) cell culture, scaffolds provide an adhesive substrate that also serves as a physical support matrix for cell culture in vitro. The present work aimed to investigate the patterns of adherence and proliferation of human, canine and murine ADSC on these three types of biomaterials. Adipose tissue was obtained from patients undergoing elective liposuction, adult C57BL/6 mice and canine healthy donors. ADSC were isolated with use of collagenase, cultured and characterized. Cells between passages 4 and 7 were associated with 3D scaffolds of calcium phosphate, collagen and OPLA (BD Biosciences), dry or pre-coated with medium with 10% fetal calf serum. Different cell concentrations were used: 10~4 and 5×10~4 cells/scaffold. Cell adherence was evaluated by counting non-adhered cells stained with Giemsa. The proliferation of ADSC in 2D and 3D conditions after 3-day incubation was determined using the MTT test. The cells adhered best to dry scaffolds, and the concentration of 5×10~4 cells/scaffold was the most adequate for adherence and proliferation. Human ADSC proliferated more rapidly on 2D than in 3D conditions, contrary to murine and canine cells. These results show that cultivation of ADSC from different species in degradable biomaterials is feasible and may be exploited for the therapeutic use of these compounds.