(393b) Expansion and Neural Differentiation of Embryonic Stem Cells (ESCs) in 3-Dimensional (3-D) Bioreactor Culture Systems

3-D bioreactor culture systems for integrated expansion and neural differentiation of ESCs were developed using polyethylene terephthalate fibrous matrices as cell supports. The fibrous matrices provided large surface areas and an in vivo-like 3-D environment, facilitating high density cell growth and regulating cell proliferation and differentiation. In this study, murine ESCs were inoculated either within small patches of PET matrices in a 125-mL bioreactor or into a fibrous bed bioreactor packed with PET matrices. The cultures were maintained in an expansion medium for 4 days, replaced with EB formation medium for 4 days, followed by astrocyte-conditioned medium to induce neural differentiation for 14 days. Cells were expanded over 20 times with a significant fraction of cells exhibiting neurogenesis as confirmed using immunostaining, flow cytometry analysis and reverse transcriptase polymerase chain reaction in both cultures. The simplified, scalable, 3-dimensional bioreactor culture systems show great potentials in producing large amounts of ES derivative cells for clinical applications.