Abstract
iPSCs are potential cell types that can be used for regenerative medicine. Suspension culture is the main approach to produce a sufficient amount of required cells to realize the application of hiPSCs for the transplantation of the large organ. The dialysis culture system holds the potential to reduce the cost by utilizing endogenous growth factors and recycle the remaining exogenous growth factors at the same time. However, the current large scale dialysis culture system was not optimized for expanding the high-density culture. Several problems such as the requirement of the large volume and technical complexity make the optimization of this system remain challenging. Also, the interference of mechanical stress in the dynamic suspension culture may reduce cellular viability, pluripotency, and differentiation capacity. Here, we describe the simple miniaturized dialysis platform to evaluate the feasibility of high-density hiPSCs culture by combining the utilization of endogenous growth factors supported by a continuous exchange of nutrition and toxic metabolic product in a low mechanical stress culture environment in a viscoelastic medium.
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References
Torizal FG, Horiguchi I, Sakai Y (2019) Physiological microenvironmental conditions in different scalable culture systems for pluripotent stem cell expansion and differentiation. Open Biomed Eng J 13:41–54
Horiguchi I, Urabe Y, Kimura K et al (2018) Effects of glucose, lactate and basic FGF as limiting factors on the expansion of human induced pluripotent stem cells. J Biosci Bioeng 125:111–115
Ouyang A, Ng R, Yang S-T (2006) Long-term culturing of undifferentiated embryonic stem cells in conditioned media and three-dimensional fibrous matrices without extracellular matrix coating. Stem Cells 25:47–454
Wolfe RP, Ahsan T (2014) Shear stress during early ESC differentiation promotes hematopoietic and endothelial phenotypes. HHS Public Access 110:1231–1242
Kehoe DE, Jing D, Lock LT et al (2010) Scalable stirred-suspension bioreactor culture. Tissue Eng 16:405–421
Kuo HH, Gao X, DeKeyser J et al (2020) Negligible-cost and weekend-free chemically defined human iPSC culture. Stem Cell Rep 14:256–270
Yamaji H, Amos B, Emery AN et al (2002) Dialysis perfusion culture of animal cells for the production of biologicals. Anim Cell Technol Basic Appl Asp 12:115–119
Amos B, Al-Rubeai M, Emery AN (1994) Hybridoma growth and monoclonal antibody production in a dialysis perfusion system. Enzym Microb Technol 16:688–695
Kurosawa H, Märkl H, Niebuhr-Redder C et al (1991) Dialysis bioreactor with radial-flow fixed bed for animal cell culture. J Ferment Bioeng 72:41–45
Shinohara M, Choi H, Ibuki M et al (2019) Endodermal differentiation of human induced pluripotent stem cells using simple dialysis culture system in suspension culture. Regen Ther 12:14–19
Griffiths B (1990) Perfusion system for cell cultivation. In: Lubiniecki A, Dekker M (eds) Mammalian cell culture technology, vol 17. Marcel Dekker, New York, pp 217–250
Côme J, Nissan X, Aubry L et al (2008) Improvement of culture conditions of human embryoid bodies using a controlled perfused and dialyzed bioreactor system. Tissue Eng Part C Methods 14:289–298
Nath SC, Nagamori E, Horie M et al (2017) Culture medium refinement by dialysis for the expansion of human induced pluripotent stem cells in suspension culture. Bioprocess Biosyst Eng 40:123–131
Xu C, Rosler E, Jiang J et al (2005) Basic fibroblast growth factor supports undifferentiated human embryonic stem cell growth without conditioned medium. Stem Cells 23:315–323
Wu J, Rostami MR, Cadavid Olaya DP et al (2014) Oxygen transport and stem cell aggregation in stirred-suspension bioreactor cultures. PLoS One 9:1–12
Otsuji TG, Bin J, Yoshimura A et al (2014) A 3D sphere culture system containing functional polymers for large-scale human pluripotent stem cell production. Stem Cell Rep 2:734–745
Acknowledgments
This research was supported by the Research Center Network for Realization of Regenerative Medicine, Japan Agency for Medical Research and Development (AMED). The FP003 solution kit was kindly provided by Nissan Chemical Corporation, Japan.
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Torizal, F.G., Choi, H., Shinohara, M., Sakai, Y. (2021). Efficient High-Density hiPSCs Expansion in Simple Dialysis Device. In: Nagy, A., Turksen, K. (eds) Induced Pluripotent Stem (iPS) Cells. Methods in Molecular Biology, vol 2454. Humana, New York, NY. https://doi.org/10.1007/7651_2021_391
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DOI: https://doi.org/10.1007/7651_2021_391
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