Abstract
Defined pluripotent stem cell culture media is a valuable tool for tracking and analyzing morphological, cell signaling and gene expression changes in human embryonic stem cells. Cultivation of hESC under xeno-free culture settings provides researchers with a consistent and reproducible environment to test experimental hypotheses and move towards translational and clinical research (Richards et al., Nat Biotechnol 20:933–936, 2002; Richards et al., Stem Cells 21:546–556, 2003). One of the primary concerns of the xenogeneic culture is the transfer of foreign pathogens or antigens that induce disease or immune response by the patient. These undesirable by-products may come from the use of murine-derived feeder cells, xenogeneic matrices, or from animal-derived components found in the cell culture medium or matrix used to isolate or expand the stem cells (Beattie et al., Stem Cells 23:489–495, 2005; Koivisto et al., Reprod Biomed Online 9:330–337, 2004). This chapter describes standardized protocols for obtaining consistent and reproducible results for culturing PSC under xeno-free, feeder-free, or feeder-based systems.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Amit M, Shariki C, Margulets V, Itskovitz-Eldor J (2004) Feeder layer- and serum-free culture of human embryonic stem cells. Biol Reprod 70:837–845
Thomson JA, Itskovitz-Eldor J, Shapiro SS, Waknitz MA, Swiergiel JJ, Marshall VS, Jones JM (1998) Embryonic stem cell lines derived from human blastocysts. Science 282:1145–1147
Rajala K, Hakala H, Panula S, Aivio S, Pihlajamaki H, Suuronen R et al (2007) Testing of nine different xeno-free culture media for human embryonic stem cell cultures. Hum Reprod 22:1231–1238
Cote RJ (2001) Aseptic technique for cell culture. Curr Protoc Cell Biol Chapter 1: Unit 1.3
Phelan MC (2007) Basic techniques in mammalian cell tissue culture. Curr Protoc Cell Biol 36:1.1.1–1.1.18
Acknowledgements
This work was supported by Life Technologies. The authors thank Drs. Pauline Lieu and Soojung Shin for their feedback and guidance.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2013 Springer Science+Business Media New York
About this protocol
Cite this protocol
Kaur, J., Tilkins, M.L., Eckert, R., Boucher, S. (2013). Methods for Culturing Human Embryonic Stem Cells in a Xeno-Free System. In: Lakshmipathy, U., Vemuri, M. (eds) Pluripotent Stem Cells. Methods in Molecular Biology, vol 997. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-348-0_9
Download citation
DOI: https://doi.org/10.1007/978-1-62703-348-0_9
Published:
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-62703-347-3
Online ISBN: 978-1-62703-348-0
eBook Packages: Springer Protocols