Abstract
The ability of human pluripotent stem cells (hPSCs) to proliferate indefinitely in culture while maintaining their pluripotent properties makes them a powerful tool for use in research, and provides tremendous potential for diagnostic testing, and therapeutic application. Success in these areas, however, is dependent on the ability to effectively expand them in long-term culture while preserving their distinct nature. Contained in this chapter are detailed protocols for the feeder-independent culture and expansion of hPSCs using mTeSR1 medium and Matrigel matrix, and guidelines for the successful transfer of those cells to alternative platforms. These protocols have been used widely by laboratories around the world to successfully expand hPSCs for long-term culture while maintaining their undifferentiated, pluripotent state.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Thomson JA, Itskovitz-Eldor J, Shapiro SS et al (1998) Embryonic stem cell lines derived from human blastocysts. Science 282:1145–1147
Reubinoff BE, Pera MF, Fong CY et al (2000) Embryonic stem cell lines from human blastocysts: somatic differentiation in vitro. Nat Biotechnol 18:399–404
Amit M, Carpenter MK, Inokuma MS et al (2000) Clonally derived human embryonic stem cell lines maintain pluripotency and proliferative potential for prolonged periods of culture. Dev Biol 227:271–278
Ludwig TE, Levenstein ME, Jones JM et al (2006) Derivation of human embryonic stem cells in defined conditions. Nat Biotechnol 24(2):185–187
Ludwig TE, Bergendahl V, Levenstein ME et al (2006) Feeder-independent culture of human embryonic stem cells. Nat Methods 3(8):637–646
International Stem Cell Initiative Consortium (2010) Comparison of defined culture systems for feeder free propagation of human embryonic stem cells. In Vitro Cell Dev Biol Anim 46(3–4):247–258
Draper JS, Moore HD, Ruban LN et al (2004) Culture and characterization of human embryonic stem cells. Stem Cells Dev 13:325–236
Buzzard JJ, Gough NM, Crook JM, Colman A (2004) Karyotype of human ES cells during extended culture. Nat Biotechnol 22:381–382
Mitalipova MM, Rao RR, Hoyer DM et al (2004) Preserving the genetic integrity of human embryonic stem cells. Nat Biotechnol 23:19–20
Xu C, Inokuma MS, Denham J et al (2001) Feeder-free growth of undifferentiated human embryonic stem cells. Nat Biotechnol 19:971–974
Chen G, Gulbranson DR, Hou Z et al (2011) Chemically defined conditions for human iPSC derivation and culture. Nat Methods 8(5):424–429
Ludwig TE, Thomson JA (2007) Defined, feeder-independent medium for human embryonic stem cell culture. Curr Protoc Stem Cell Biol:1C.2.1–1C.2.16
Braam SR, Zeinstra L, Litjens S et al (2008) Recombinant vitronectin is a functionally defined substrate that supports human embryonic stem cell self-renewal via alphavbeta5 integrin. Stem Cells 26(9):2257–2265
Fazeli A, Liew CG, Matin MM et al (2011) Altered patterns of differentiation in karyotypically abnormal human embryonic stem cells. Int J Dev Biol 55(2):175–180
Acknowledgment
This work was supported by funding from the Wisconsin Alumni Research Foundation.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2017 Springer Science+Business Media LLC
About this protocol
Cite this protocol
Brehm, J.L., Ludwig, T.E. (2017). Culture, Adaptation, and Expansion of Pluripotent Stem Cells. In: Crook, J., Ludwig, T. (eds) Stem Cell Banking. Methods in Molecular Biology, vol 1590. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6921-0_10
Download citation
DOI: https://doi.org/10.1007/978-1-4939-6921-0_10
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-6919-7
Online ISBN: 978-1-4939-6921-0
eBook Packages: Springer Protocols