Category: Cellular Technologies

1113-D - Use of Hydrodynamic Force in Stirred Suspension Culture of Human Pluripotent Stem Cells to Regulate Expansion and Differentiation

Tuesday, February 6, 2018
5:00 PM - 6:00 PM

Use of Hydrodynamic Force in Stirred Suspension Culture of Human Pluripotent Stem Cells to Regulate Expansion and Differentiation
Hideaki Tsutsui, Ph.D.
University of California, Riverside
Riverside, CA USA

Daniel Nampe, Ronak Joshi, Joshua Karam, Kevin Keller, Nicole I. zur Nieden

Human pluripotent stem cells (hPSCs) are a promising cell source for the future stem cell therapy and regenerative medicine. Many such target applications require a billion or more stem cells and their derivatives to treat each patient. Conventional laboratory-scale, 2-D adherent cultures are not efficient for producing such a large number of cells in a cost-effective manner. Three-dimensional (3-D) stirred suspension culture has been adapted for the hPSCs in the last several years. This new approach, however, requires optimization for hPSCs, as they are very sensitive to external physical stimuli, unlike Chinese hamster ovary cells and hybridomas that are robust and to which the conventional stirred suspension cultures have been tailored. As a result, the most common strategy in stem cell stirred suspension culture is to use weak mixing that is adequate for achieving chemical homogeneity while preventing damages to the cells. Our approach, on the other hand, contrasts with those by actively using fluidic forces to regulate the size of growing cell aggregates and even intervening possible cellular mechanotransduction mechanisms to our advantage. We show that an optimal fluidic agitation maintains the cell aggregate below their critical size beyond which the cells suffer from spontaneous differentiation and even apoptosis due to the diffusion limit. As a result, the cells under this condition can expand 38 fold over 7 days of culture, approximately twice and 4 times faster than the conventional 2-D and 3-D static cultures. These results were accompanied by an interesting Western blot data, clearly showing activation/deactivation of known cell-signaling kinases that are dependent on the intensity of fluidic agitation. This discovery was leveraged to enhance differentiation of hPSCs into cardiomyocytes using a stage-specific chemical induction protocol in conjunction with dynamic fluidic agitation inputs. Although this part of the work needs more in-depth investigations, our preliminary results show that the use of fluidic force can facilitate maturation of hPSC-derived cardiomyocytes significantly, where distinct signature of mature phenotype was observed within 25 days of cardiac induction. This approach is expected to pave a way for the development of an innovative stem cell biomanufacturing platform.

Hideaki Tsutsui

Assistant Professor
University of California, Riverside
Riverside, CA

Hideaki Tsutsui is an assistant professor of mechanical engineering at the University of California, Riverside.