Abstract
Cell culture media are often supplemented with recombinant growth factors and cytokines to reproduce biological conditions in vitro. Basic fibroblast growth factor (bFGF) has been widely used to support the pluripotency and self-renewal activity of human induced pluripotent stem cells (hiPSCs). We had previously developed a synthetic surrogate for bFGF on the basis of a DNA aptamer that binds to one of the FGF receptors. Since DNA aptamers have advantages over recombinant proteins in terms of thermal stability and production cost, replacing recombinant growth factors in cell culture media with DNA aptamers would be of great interest. Herein, we describe our protocol for feeder-free hiPSC culture using a DNA aptamer-based mimic of bFGF.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Keefe AD, Pai S, Ellington A (2010) Aptamers as therapeutics. Nat Rev Drug Discov 9:537–550
Zhou J, Rossi J (2017) Aptamers as targeted therapeutics: current potential and challenges. Nat Rev Drug Discov 16:181–202
Ueki R, Ueki A, Kanda N, Sando S (2016) Oligonucleotide-based mimetics of hepatocyte growth factor. Angew Chem Int Ed 55:579–582
Ueki R, Atsuta S, Ueki A, Sando S (2017) Nongenetic reprogramming of the ligand specificity of growth factor receptors by bispecific DNA aptamers. J Am Chem Soc 139:6554–6557
Ueki R, Atsuta S, Ueki A, Hoshiyama J, Li J, Hayashi Y, Sando S (2019) DNA aptamer assemblies as fibroblast growth factor mimics and their application in stem cell culture. Chem Commun 55:2672–2675
Kamatkar N, Levy M, Hébert JM (2019) Development of a monomeric inhibitory RNA aptamer specific for FGFR3 that acts as an activator when dimerized. Mol Ther Nucleic Acids 17:530–539
Ramaswamy V, Monsalve A, Sautina L, Segal MS, Dobson J, Allen JB (2015) DNA aptamer assembly as a vascular endothelial growth factor receptor agonist. Nucleic Acid Ther 25:227–234
Yoshitomi T, Hayashi M, Oguro T, Kimura K, Wayama F, Furusho H, Yoshimoto K (2020) Binding and structural properties of DNA aptamers with VEGF-A-mimic activity. Mol Ther Nucleic Acids 19:1145–1152
Miyazaki T, Isobe T, Nakatsuji N, Suemori H (2017) Efficient adhesion culture of human pluripotent stem cells using laminin fragments in an uncoated manner. Sci Rep 7:41165
Acknowledgments
This work was supported by a Leave a Nest Scientific Research grant from Ikedarika Scientific Co., Ltd. and Leave a Nest Co., Ltd., and a Noguchi-Shitagau Research Grant from the Noguchi Institute, and partly by a research grant from Asahi Glass Foundation.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2021 Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Hayata, Y., Ueki, R., Sando, S. (2021). Feeder-Free Human Induced Pluripotent Stem Cell Culture Using a DNA Aptamer-Based Mimic of Basic Fibroblast Growth Factor. In: Kojima, R. (eds) Mammalian Cell Engineering. Methods in Molecular Biology, vol 2312. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1441-9_18
Download citation
DOI: https://doi.org/10.1007/978-1-0716-1441-9_18
Published:
Publisher Name: Humana, New York, NY
Print ISBN: 978-1-0716-1440-2
Online ISBN: 978-1-0716-1441-9
eBook Packages: Springer Protocols