title: Optimization of agitation speed in spinner flask for microcarrier structural integrity and expansion of induced pluripotent stem cells.
abstract: In recent times, the study and use of induced pluripotent stem cells (iPSC) have become important in order to avoid the ethical issues surrounding the use of embryonic stem cells. Therapeutic, industrial and research based use of iPSC requires large quantities of cells generated in vitro. Mammalian cells, including pluripotent stem cells, have been expanded using 3D culture, however current limitations have not been overcome to allow a uniform, optimized platform for dynamic culture of pluripotent stem cells to be achieved. In the current work, we have expanded mouse iPSC in a spinner flask using Cytodex 3 microcarriers. We have looked at the effect of agitation on the microcarrier survival and optimized an agitation speed that supports bead suspension and iPS cell expansion without any bead breakage. Under the optimized conditions, the mouse iPSC were able to maintain their growth, pluripotency and differentiation capability. We demonstrate that microcarrier survival and iPS cell expansion in a spinner flask are reliant on a very narrow range of spin rates, highlighting the need for precise control of such set ups and the need for improved design of more robust systems.
authors: IITB Monash Research Academy, Mumbai, India. priyankka.g@gmail.com.|Department of Chemical Engineering, IIT Bombay, Mumbai, India. priyankka.g@gmail.com.|Department of Chemical Engineering, Monash University, Melbourne, VIC, Australia. priyankka.g@gmail.com.|Division of Biological Engineering, Monash University, Melbourne, VIC, Australia. priyankka.g@gmail.com.
authors: Priyanka
authors: P
authors: Gupta
authors: Division of Biological Engineering, Monash University, Melbourne, VIC, Australia.|Department of Mechanical and Aerospace Engineering, Monash University, Melbourne, VIC, Australia.
authors: Mohd-Zulhilmi
authors: MZ
authors: Ismadi
authors: Division of Biological Engineering, Monash University, Melbourne, VIC, Australia.|South Australian Research and Development Institute, Rosedale, SA, Australia.
authors: Paul J
authors: PJ
authors: Verma
authors: Department of Mechanical and Aerospace Engineering, Monash University, Melbourne, VIC, Australia.
authors: Andreas
authors: A
authors: Fouras
authors: Department of Chemical Engineering, IIT Bombay, Mumbai, India.
authors: Sameer
authors: S
authors: Jadhav
authors: Department of Chemical Engineering, IIT Bombay, Mumbai, India.
authors: Jayesh
authors: J
authors: Bellare
authors: Division of Biological Engineering, Monash University, Melbourne, VIC, Australia.|Department of Mechanical and Aerospace Engineering, Monash University, Melbourne, VIC, Australia.
authors: Kerry
authors: K
authors: Hourigan
keywords: Induced pluripotent stem cells; Microcarrier; Spinner flask