(538c) Bioreactor and Media Development for Amniotic Fluid Stem Cells

Human amniotic fluid stem cells (hAFSCs) are emerging as an important cell source for tissue engineering and regenerative medicine due to their easy accessibility and broad multi-potentiality. In clinical applications, a large number of hAFSCs and chemically defined media are required. To address these issues, the expansion of hAFSCs in 3-dimensional (3-D) polyethylene terephthalate (PET) scaffolds in a stirred bioreactor, and the effect of currculigoside and naringin as potential components of chemical defined media on osteogenic differentiation of AFSCs were evaluated. The results showed that 3-D PET scaffold with in vivo-like environment and a large specific surface area for cell adhesion promoted cell expansion (66-fold vs. 38-fold) compared to 2-D culture. A dynamic fibrous bed bioreactor (FBB) was used to expand hAFSCs to reach a high cell density of 3.2 × 10⁶ cells/mL. The bioreactor-expanded cells maintained clonogenic ability, high levels of expression of characteristic stem cell surface makers, and very good multi-linage differentiation capabilities.  Curculigoside and naringin stimulated alkaline phosphatase activity, calcium deposition, and expression of the osteogenic genes of hAFSCs during osteogenic differentiation in a dose-dependent manner (1–100 mg/mL). Concurrently, osteoclastogenesis of hAFSCs were inhibited by curculigoside. Moreover, the roles of Wnt/β -catenin and BMP signaling pathways were revealed during curculigoside and naringin treatments, respectively. In summary, this study demonstrated the feasibility of using the FBB to mass-produce hAFSCs, and the potential use of curculigoside and naringin for chemically defined media to induce hAFSC osteogenic differentiation for future applications in tissue engineering and regenerative medicine.