| With the growing interest towards the use of pluripotent embryonic stem cells (ESCs) in tissue engineering and cell therapies, the current study was undertaken to develop a controlled and reproducible process for the large-scale production of ESCs. Protocols were developed to successfully expand murine ESCs in suspension bioreactors and, through serial passaging in suspension, a substantial increase in the expansion of the cells was achieved over an extended culture period. Importantly, pluripotency of the cells was maintained, as determined through protein and gene expression of pluripotency and early differentiation markers, colony-forming efficiency and multilineage differentiation. Further, effective bioreactor protocols were developed for embryoid body (EB) formation, and preliminary results supported that the EBs could undergo induced differentiation in suspension. With the development of effective bioreactor protocols for ESC expansion and differentiation, the current study has overcome a major obstacle in the progress towards the clinical implementation of human ESC technology. |
